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1.
J Infect Dev Ctries ; 14(10): 1138-1145, 2020 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-33175709

RESUMO

INTRODUCTION: The coronavirus disease 2019 (COVID-19) pandemic is spreading rapidly. Critically ill cases of COVID-19 can rapidly progress to acute respiratory distress syndrome and multiple organ failures. However, no effective drugs have been available till now, leading to more than 300,000 deaths up to 29 April 2020. Here, we present a critically ill case utilizing umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs). CASE PRESENTATION: A 72-year-old man was admitted, with the diagnosis of COVID-19, ARDS, type-2 diabetes, diabetic nephropathy, renal insufficiency, and hypertension. His clinical condition continually developed to be life-threatening even receiving various treatment options including antiviral therapy and extracorporeal membrane oxygenation. Between 28 February and 8 March 2020, the patient was given 5-time intravenous infusions of UCB-MSCs. His hematological and biochemical indexes, including lymphocytes and renal function improved. Pulmonary static compliance increased significantly and PaO2/FiO2 ratio maintained stable. On March 10, he received lung transplantation. CONCLUSIONS: Our current findings suggested that UCB-MSCs therapy may show some positive effect in treating critical COVID-19 to some extent, for its delaying deterioration of the disease and efficacy in respiratory and renal function, though limited.


Assuntos
Infecções por Coronavirus/terapia , Sangue Fetal/citologia , Transplante de Células-Tronco Mesenquimais , Pneumonia Viral/terapia , Idoso , Betacoronavirus , Estado Terminal , Evolução Fatal , Humanos , Transplante de Pulmão , Masculino , Pandemias
2.
PLoS One ; 15(9): e0239495, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32956417

RESUMO

Cell-type specific gene expression profiles are needed for many computational methods operating on bulk RNA-Seq samples, such as deconvolution of cell-type fractions and digital cytometry. However, the gene expression profile of a cell type can vary substantially due to both technical factors and biological differences in cell state and surroundings, reducing the efficacy of such methods. Here, we investigated which factors contribute most to this variation. We evaluated different normalization methods, quantified the variance explained by different factors, evaluated the effect on deconvolution of cell type fractions, and examined the differences between UMI-based single-cell RNA-Seq and bulk RNA-Seq. We investigated a collection of publicly available bulk and single-cell RNA-Seq datasets containing B and T cells, and found that the technical variation across laboratories is substantial, even for genes specifically selected for deconvolution, and this variation has a confounding effect on deconvolution. Tissue of origin is also a substantial factor, highlighting the challenge of using cell type profiles derived from blood with mixtures from other tissues. We also show that much of the differences between UMI-based single-cell and bulk RNA-Seq methods can be explained by the number of read duplicates per mRNA molecule in the single-cell sample. Our work shows the importance of either matching or correcting for technical factors when creating cell-type specific gene expression profiles that are to be used together with bulk samples.


Assuntos
Linfócitos B/química , Análise de Sequência de RNA , Linfócitos T/química , Transcriptoma , Adulto , Composição de Bases , Conjuntos de Dados como Assunto , Sangue Fetal/citologia , Humanos , Recém-Nascido , Análise de Componente Principal , Análise de Célula Única , Manejo de Espécimes
3.
Exp Parasitol ; 217: 107938, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32768560

RESUMO

PURPOSE: Praziquantel (PZQ) is the conventional antibilharzial agent. Nevertheless, no antibilharzial prophylactic agents or 100% curable therapy approved and no reported data about use of human CD34+ Umbilical Cord Blood Stem Cells (CD34+UCBSCs) or Wharton Jelly Mesenchymal Stem Cells (WJMSCs) in prevention and/or complete eradication of acute S.mansoni granulomas in liver. We aimed to study possible prophylactic vs therapeutic role of human CD34+UCBSCs and WJMSCs in acute hepatic bilharzial granulomas in pre vs post-infected mice. METHODS: Seventy mice were divided into 7 groups (10 mice each): Normal, S.mansoni-infected, post-infected PZQ-treated, CD34+UCBSCs pre and post-infected, WJMSCs pre and post-infected. Serological, parasitological, histopathological evaluation using OCT4 & TGFB immunohistochemistry and quantitative image analysis assessment of TGFB-stained fibrogenesis in liver granulomas performed. RESULTS: Histopathologically, surprisingly and significantly, the prophylactic pre-infection stem cells (CD34+UCBSCs and WJMSCs) & similarly the post-infection CD34+UCBSCs treatment revealed eradication/reversal of the entire granulomas and no fibrosis. Moreover, post-infection PZQ treatment showed fewer and significantly smaller granulomas than post-infection WJMSCs treatment. Nevertheless, post-infection WJMSCs exhibited non-significant less TGFB-stained fibrogenesis. CONCLUSION: CD34+UCBSCs exerted the best prophylactic and therapeutic roles in prevention and complete cure of acute hepatic S.mansoni granulomas over WJMSCs and PZQ. In contrast, only pre-infection WJMSCs exhibited similar preventive (prophylactic) effect. On the contrary, post-infection WJMSCs were the worst (incompletely reversed granulomas). Post-infection Praziquantel was overall better therapeutically than WJMSCs in this regard. Accordingly, when it comes to WJMSCs application, WJMSCs are better used as a pre-infection prophylactic and preventive tool rather than a post-infection therapy. Further studies are needed.


Assuntos
Antígenos CD34/sangue , Transplante de Células-Tronco de Sangue do Cordão Umbilical , Esquistossomose mansoni/prevenção & controle , Esquistossomose mansoni/terapia , Animais , Anti-Helmínticos/administração & dosagem , Fezes/parasitologia , Sangue Fetal/citologia , Citometria de Fluxo , Granuloma/prevenção & controle , Granuloma/terapia , Humanos , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica , Fígado/patologia , Hepatopatias Parasitárias/prevenção & controle , Hepatopatias Parasitárias/terapia , Masculino , Células-Tronco Mesenquimais , Camundongos , Fator 3 de Transcrição de Octâmero , Contagem de Ovos de Parasitas , Praziquantel/administração & dosagem , Coloração e Rotulagem , Fator de Crescimento Transformador beta
4.
Cochrane Database Syst Rev ; 8: CD013202, 2020 08 19.
Artigo em Inglês | MEDLINE | ID: mdl-32813884

RESUMO

BACKGROUND: Hypoxic-ischaemic encephalopathy (HIE) is a leading cause of mortality and long-term neurological sequelae, affecting thousands of children worldwide. Current therapies to treat HIE are limited to cooling. Stem cell-based therapies offer a potential therapeutic approach to repair or regenerate injured brain tissue. These preclinical findings have now culminated in ongoing human neonatal trials. OBJECTIVES: To determine the efficacy and safety of stem cell-based interventions for the treatment of hypoxic-ischaemic encephalopathy (HIE) in newborn infants. SEARCH METHODS: We used the standard search strategy of Cochrane Neonatal to search the Cochrane Central Register of Controlled Trials (CENTRAL; 2020, Issue 5), MEDLINE via PubMed (1966 to 8 June 2020), Embase (1980 to 8 June 2020), and CINAHL (1982 to 8 June 2020). We also searched clinical trials databases, conference proceedings, and the reference lists of retrieved articles for randomised controlled trials and quasi-randomised trials. SELECTION CRITERIA: Randomised controlled trials, quasi-randomised controlled trials and cluster trials comparing 1) stem cell-based interventions (any type) compared to control (placebo or no treatment); 2) use of mesenchymal stem/stromal cells (MSCs) of type (e.g. number of doses or passages) or source (e.g. autologous versus allogeneic, or bone marrow versus cord) versus MSCs of other type or source; 3) use of stem cell-based interventions other than MSCs of type (e.g. mononuclear cells, oligodendrocyte progenitor cells, neural stem cells, hematopoietic stem cells, and inducible pluripotent stem cells) or source (e.g. autologous versus allogeneic, or bone marrow versus cord) versus stem cell-based interventions other than MSCs of other type or source; or 4) MSCs versus stem cell-based interventions other than MSCs. DATA COLLECTION AND ANALYSIS: For each of the included trials, two authors independently planned to extract data (e.g. number of participants, birth weight, gestational age, type and source of MSCs or other stem cell-based interventions) and assess the risk of bias (e.g. adequacy of randomisation, blinding, completeness of follow-up). The primary outcomes considered in this review are all-cause neonatal mortality, major neurodevelopmental disability, death or major neurodevelopmental disability assessed at 18 to 24 months of age. We planned to use the GRADE approach to assess the quality of evidence. MAIN RESULTS: Our search strategy yielded 616 references. Two review authors independently assessed all references for inclusion. We did not find any completed studies for inclusion. Fifteen RCTs are currently registered and ongoing. We describe the three studies we excluded. AUTHORS' CONCLUSIONS: There is currently no evidence from randomised trials that assesses the benefit or harms of stem cell-based interventions for the prevention of morbidity and mortality following hypoxic-ischaemic encephalopathy in newborn infants.


Assuntos
Hipóxia-Isquemia Encefálica/terapia , Transplante de Células-Tronco , Anti-Inflamatórios/uso terapêutico , Sangue Fetal/citologia , Humanos , Hidrocortisona/uso terapêutico , Hipotermia Induzida , Hipóxia-Isquemia Encefálica/complicações , Hipóxia-Isquemia Encefálica/mortalidade , Recém-Nascido
5.
Nat Commun ; 11(1): 3761, 2020 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-32724101

RESUMO

Chronic immune-mediated diseases of adulthood often originate in early childhood. To investigate genetic associations between neonatal immunity and disease, we map expression quantitative trait loci (eQTLs) in resting myeloid cells and CD4+ T cells from cord blood samples, as well as in response to lipopolysaccharide (LPS) or phytohemagglutinin (PHA) stimulation, respectively. Cis-eQTLs are largely specific to cell type or stimulation, and 31% and 52% of genes with cis-eQTLs have response eQTLs (reQTLs) in myeloid cells and T cells, respectively. We identified cis regulatory factors acting as mediators of trans effects. There is extensive colocalisation between condition-specific neonatal cis-eQTLs and variants associated with immune-mediated diseases, in particular CTSH had widespread colocalisation across diseases. Mendelian randomisation shows causal neonatal gene expression effects on disease risk for BTN3A2, HLA-C and others. Our study elucidates the genetics of gene expression in neonatal immune cells, and aetiological origins of autoimmune and allergic diseases.


Assuntos
Doenças Autoimunes/genética , Desenvolvimento Infantil/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/imunologia , Hipersensibilidade/genética , Locos de Características Quantitativas/imunologia , Doenças Autoimunes/imunologia , Butirofilinas/genética , Butirofilinas/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Catepsina H/genética , Catepsina H/metabolismo , Criança , Pré-Escolar , Conjuntos de Dados como Assunto , Sangue Fetal/citologia , Perfilação da Expressão Gênica , Redes Reguladoras de Genes/imunologia , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Antígenos HLA-C/genética , Antígenos HLA-C/metabolismo , Humanos , Hipersensibilidade/imunologia , Lactente , Recém-Nascido , Análise da Randomização Mendeliana , Células Mieloides/imunologia , Células Mieloides/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Polimorfismo de Nucleotídeo Único , Estudos Prospectivos
6.
Proc Natl Acad Sci U S A ; 117(23): 12868-12876, 2020 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-32457162

RESUMO

Fine-resolution differentiation trajectories of adult human hematopoietic stem cells (HSCs) involved in the generation of red cells is critical for understanding dynamic developmental changes that accompany human erythropoiesis. Using single-cell RNA sequencing (scRNA-seq) of primary human terminal erythroid cells (CD34-CD235a+) isolated directly from adult bone marrow (BM) and umbilical cord blood (UCB), we documented the transcriptome of terminally differentiated human erythroblasts at unprecedented resolution. The insights enabled us to distinguish polychromatic erythroblasts (PolyEs) at the early and late stages of development as well as the different development stages of orthochromatic erythroblasts (OrthoEs). We further identified a set of putative regulators of terminal erythroid differentiation and functionally validated three of the identified genes, AKAP8L, TERF2IP, and RNF10, by monitoring cell differentiation and apoptosis. We documented that knockdown of AKAP8L suppressed the commitment of HSCs to erythroid lineage and cell proliferation and delayed differentiation of colony-forming unit-erythroid (CFU-E) to the proerythroblast stage (ProE). In contrast, the knockdown of TERF2IP and RNF10 delayed differentiation of PolyE to OrthoE stage. Taken together, the convergence and divergence of the transcriptional continuums at single-cell resolution underscore the transcriptional regulatory networks that underlie human fetal and adult terminal erythroid differentiation.


Assuntos
Diferenciação Celular/genética , Eritroblastos/fisiologia , Eritropoese/genética , Adulto , Apoptose/genética , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Sangue Fetal/citologia , Técnicas de Silenciamento de Genes , Redes Reguladoras de Genes , Humanos , Recém-Nascido , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Masculino , Família Multigênica , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , RNA-Seq , Análise de Célula Única , Proteínas de Ligação a Telômeros/genética , Proteínas de Ligação a Telômeros/metabolismo , Transcrição Genética
7.
Hum Cell ; 33(3): 470-475, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32447573

RESUMO

Osteoarthritis is a chronic degenerative joint disease with an incidence of 81% among people aged over 65 years in China. Osteoarthritis significantly decreases the quality of life of patients, causing physical and psychological damage and posing a serious economic burden. Clinical treatments for osteoarthritis include drug and surgical treatments. Drug treatment can successfully alleviate pain but not satisfactorily reverse joint damage, while surgical intervention is typically used to treat end-stage disease. Stem cells are multi-potential progenitor cells with self-renewal and multi-lineage differentiation abilities, and can differentiate into many kinds of cells, including chondrocytes. Umbilical cord stem cells, also known as Wharton's jelly mesenchymal stem cells (WJ-MSCs), have become the first choice for cartilage regeneration engineering owing to their availability and convenience of collection. This article reviews the biological characterization of WJ-MSCs in recent years, their advantages compared with other stem cells, and their application in the treatment of osteoarthritis in animal experiments and clinical trials.


Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Osteoartrite/terapia , Animais , Diferenciação Celular , Células Cultivadas , Sangue Fetal/citologia , Humanos , Células-Tronco Mesenquimais/imunologia , Células-Tronco Mesenquimais/fisiologia
8.
Proc Natl Acad Sci U S A ; 117(16): 8845-8849, 2020 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-32253306

RESUMO

The genetic incorporation of noncanonical amino acids (ncAAs) into proteins has been realized in bacteria, yeast, and mammalian cells, and recently, in multicellular organisms including plants and animals. However, the addition of new building blocks to the genetic code of tissues from human origin has not yet been achieved. To this end, we report a self-replicating Epstein-Barr virus-based episomal vector for the long-term encoding of ncAAs in human hematopoietic stem cells and reconstitution of this genetically engineered hematopoietic system in mice.


Assuntos
Aminoácidos/genética , Diferenciação Celular/genética , Vetores Genéticos/genética , Células-Tronco Hematopoéticas/fisiologia , Engenharia de Proteínas/métodos , Animais , Sangue Fetal/citologia , Técnicas de Transferência de Genes , Código Genético , Células HEK293 , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 4/genética , Humanos , Camundongos , Camundongos Endogâmicos NOD , Plasmídeos/genética , Cultura Primária de Células/métodos , Transfecção/métodos , Quimeras de Transplante , Transplante Heterólogo/métodos
9.
Sci Rep ; 10(1): 4603, 2020 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-32165664

RESUMO

Neonatal hypoxic-ischaemic encephalopathy (HIE) is a serious condition; many survivors develop neurological impairments, including cerebral palsy and intellectual disability. Preclinical studies show that the systemic administration of umbilical cord blood cells (UCBCs) is beneficial for neonatal HIE. We conducted a single-arm clinical study to examine the feasibility and safety of intravenous infusion of autologous UCBCs for newborns with HIE. When a neonate was born with severe asphyxia, the UCB was collected, volume-reduced, and divided into three doses. The processed UCB was infused at 12-24, 36-48, and 60-72 hours after the birth. The designed enrolment was six newborns. All six newborns received UCBC therapy strictly adhering to the study protocol together with therapeutic hypothermia. The physiological parameters and peripheral blood parameters did not change much between pre- and postinfusion. There were no serious adverse events that might be related to cell therapy. At 30 days of age, the six infants survived without circulatory or respiratory support. At 18 months of age, neurofunctional development was normal without any impairment in four infants and delayed with cerebral palsy in two infants. This pilot study shows that autologous UCBC therapy is feasible and safe.


Assuntos
Transplante de Células-Tronco de Sangue do Cordão Umbilical , Sangue Fetal/citologia , Hipóxia-Isquemia Encefálica/etiologia , Hipóxia-Isquemia Encefálica/terapia , Biomarcadores , Gasometria , Terapia Baseada em Transplante de Células e Tecidos/efeitos adversos , Terapia Baseada em Transplante de Células e Tecidos/métodos , Transplante de Células-Tronco de Sangue do Cordão Umbilical/efeitos adversos , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Eletroencefalografia , Feminino , Humanos , Hipóxia-Isquemia Encefálica/diagnóstico , Hipóxia-Isquemia Encefálica/metabolismo , Recém-Nascido , Masculino , Projetos Piloto
10.
Arch Dis Child Fetal Neonatal Ed ; 105(6): 572-580, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32152192

RESUMO

OBJECTIVE: To conduct a systematic review and meta-analysis of the efficacy and safety of umbilical cord milking in preterm infants. DESIGN: Randomised controlled trials comparing umbilical cord milking with delayed cord clamping/immediate cord clamping in preterm infants were identified by searching databases, clinical trial registries and reference list of relevant studies in November 2019. Fixed effects model was used to pool the data on various clinically relevant outcomes. MAIN OUTCOME MEASURES: Mortality and morbidities in preterm neonates. RESULTS: Nineteen studies (2014 preterm infants) were included. Five studies (n=922) compared cord milking with delayed cord clamping, whereas 14 studies (n=1092) compared milking with immediate cord clamping. Cord milking, as opposed to delayed cord clamping, significantly increased the risk of intraventricular haemorrhage (grade III or more) (risk ratio (RR): 1.95 (95% CI 1.01 to 3.76), p=0.05). When compared with immediate cord clamping, cord milking reduced the need for packed RBC transfusions (RR:0.56 (95% CI 0.43 to 0.73), p<0.001). There was limited information on long-term neurodevelopmental outcomes. The grade of evidence was moderate or low for the various outcomes analysed. CONCLUSION: Umbilical cord milking, when compared with delayed cord clamping, significantly increased the risk of severe intraventricular haemorrhage in preterm infants, especially at lower gestational ages. Cord milking, when compared with immediate cord clamping, reduced the need for packed RBC transfusions but did not improve clinical outcomes. Hence, cord milking cannot be considered as placental transfusion strategy in preterm infants based on the currently available evidence.


Assuntos
Constrição , Recém-Nascido Prematuro/sangue , Assistência Perinatal/métodos , Cordão Umbilical/irrigação sanguínea , Causas de Morte , Hemorragia Cerebral/etiologia , Desenvolvimento Infantil , Parto Obstétrico/métodos , Contagem de Eritrócitos , Transfusão de Eritrócitos , Feminino , Sangue Fetal/citologia , Sangue Fetal/fisiologia , Idade Gestacional , Humanos , Lactente , Recém-Nascido , Fototerapia , Circulação Placentária , Gravidez , Fatores de Risco , Fatores de Tempo , Cordão Umbilical/fisiologia
11.
J Leukoc Biol ; 107(6): 1023-1032, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32064671

RESUMO

The Vγ9Vδ2 T cell subset is the major γδ T cell subset in human peripheral blood and has the unique ability to contribute to immune surveillance by detecting pyrophosphorylated metabolites of isoprenoid synthesis, termed phosphoantigens (pAgs). Vγ9Vδ2 T cells are first detected at midgestation and show postnatal expansion. Interestingly, neonatal Vγ9Vδ2 T cells display a higher TCR repertoire diversity with more public clonotypes and lower pAg responsiveness than in adults. Notably, it is not known whether postnatal changes occur by TCR-dependent reactivity to pAg exposure. Here, we applied next-generation sequencing of γδ TCR repertoires to understand potential differences in the pAg-mediated response of neonatal and adult Vγ9Vδ2 T cells at the level of the expressed γδ TCR. We observed a polyclonal pAg-induced response of neonatal and adult Vγ9Vδ2 T cells, albeit neonatal γδ T cells showed less in vitro pAg responsiveness. Neonatal Vγ9Vδ2 T cells displayed a less pronounced bias for Jδ1 usage and a more frequent use of Jδ2 or Jδ3 that remained stable after pAg exposure. In addition, public and private Vδ2 TRD clones took part in the polyclonal pAg-induced response in neonates and adults. In conclusion, adult and neonatal Vγ9Vδ2 T cells both undergo polyclonal pAg-induced proliferation, whereas especially adult Vγ9Vδ2 T cells display a high stability at the level of the expressed TCR repertoire.


Assuntos
Sangue Fetal/imunologia , Fosfoproteínas/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Subpopulações de Linfócitos T/imunologia , Adulto , Proliferação de Células/efeitos dos fármacos , Células Clonais , Sangue Fetal/citologia , Sangue Fetal/efeitos dos fármacos , Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Recém-Nascido , Interleucina-2/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Organofosfatos/farmacologia , Fosfoproteínas/genética , Cultura Primária de Células , Receptores de Antígenos de Linfócitos T gama-delta/genética , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/efeitos dos fármacos , Ácido Zoledrônico/farmacologia
12.
Transfusion ; 60(3): 588-597, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32056228

RESUMO

BACKGROUND: Umbilical cord blood has become an important source of hematopoietic stem and progenitor cells for therapeutic applications. However, cord blood banking (CBB) grapples with issues related to economic viability, partially due to high discard rates of cord blood units (CBUs) that lack sufficient total nucleated cells for storage or therapeutic use. Currently, there are no methods available to assess the likelihood of CBUs meeting storage criteria noninvasively at the collection site, which would improve CBB efficiency and economic viability. MATERIALS AND METHODS: To overcome this limitation, we apply a novel label-free optical imaging method, called quantitative oblique back-illumination microscopy (qOBM), which yields tomographic phase and absorption contrast to image blood inside collection bags. An automated segmentation algorithm was developed to count white blood cells and red blood cells (RBCs) and assess hematocrit. Fifteen CBUs were measured. RESULTS: qOBM clearly differentiates between RBCs and nucleated cells. The cell-counting analysis shows an average error of 13% compared to hematology analysis, with a near-perfect, one-to-one relationship (slope = 0.94) and strong correlation coefficient (r = 0.86). Preliminary results to assess hematocrit also show excellent agreement with expected values. Acquisition times to image a statistically significant number of cells per CBU were approximately 1 minute. CONCLUSION: qOBM exhibits robust performance for quantifying blood inside collection bags. Because the approach is automated and fast, it can potentially quantify CBUs within minutes of collection, without breaching the CBUs' sterile environment. qOBM can reduce costs in CBB by avoiding processing expenses of CBUs that ultimately do not meet storage criteria.


Assuntos
Sangue Fetal/citologia , Leucócitos/citologia , Microscopia/métodos , Bancos de Sangue/estatística & dados numéricos , Doadores de Sangue/estatística & dados numéricos , Coleta de Amostras Sanguíneas , Humanos
13.
F1000Res ; 92020.
Artigo em Inglês | MEDLINE | ID: mdl-31984133

RESUMO

Since the first hematopoietic stem cell transplant, over a million transplants have been performed worldwide. In the last decade, the transplant field has witnessed a progressive decline in bone marrow and cord blood utilization and a parallel increase in peripheral blood as a source of stem cells. Herein, we review the use of bone marrow and cord blood in the hematopoietic stem cell transplant setting, and we describe the recent advances made in different medical fields using cells derived from cord blood and bone marrow.


Assuntos
Medula Óssea , Terapia Baseada em Transplante de Células e Tecidos , Sangue Fetal/citologia , Transplante de Células-Tronco Hematopoéticas , Humanos
14.
Int J Mol Med ; 45(2): 556-568, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31894311

RESUMO

Alopecia is a common and distressing condition, and developing new therapeutic agents to prevent hair loss is important. Human umbilical cord blood­derived mesenchymal stem cells (hUCB­MSCs) have been studied intensively in regenerative medicine. However, the therapeutic potential of these cells against hair loss and hair organ damage remains unclear, and the effects of hUCB­MSC transplantation on hair loss require evaluation. The current study aimed to investigate the effects of hUCB­MSCs on hair regression in vivo and restoration of anagen conduction on hair growth in vitro. The effects of hUCB­MSCs were explored in mouse catagen induction models using a topical treatment of 0.1% dexamethasone to induce hair regression. Dexamethasone was also used to simulate a stress environment in vitro. The results demonstrated that hUCB­MSCs significantly prevented hair regression induced by dexamethasone topical stimulation in vivo. Additionally, hUCB­MSCs significantly increased the proliferation of human dermal papilla cells (hDPCs) and HaCaT cells, which are key constituent cells of the hair follicle. Stimulation of vascular endothelial growth factor secretion and decreased expression of DKK­1 by hUCB­MSCs were also observed in hDPCs. Restoration of cell viability by hUCB­MSCs suggested that these cells exerted a protective effect on glucocorticoid stress­associated hair loss. In addition, anti­apoptotic effects and regulation of the autophagic flux recovery were observed in HaCaT cells. The results of the present study indicated that hUCB­MSCs may have the capacity to protect hair follicular dermal papilla cells and keratinocytes, thus preventing hair loss. Additionally, the protective effects of hUCB­MSCs may be resistant to dysregulation of autophagy under harmful stress.


Assuntos
Anti-Inflamatórios/efeitos adversos , Dexametasona/efeitos adversos , Folículo Piloso/citologia , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Animais , Apoptose/efeitos dos fármacos , Linhagem Celular , Células Cultivadas , Feminino , Sangue Fetal/citologia , Cabelo/citologia , Cabelo/efeitos dos fármacos , Cabelo/crescimento & desenvolvimento , Cabelo/ultraestrutura , Folículo Piloso/efeitos dos fármacos , Folículo Piloso/ultraestrutura , Humanos , Camundongos Endogâmicos C57BL
15.
Eur J Cell Biol ; 99(2-3): 151069, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31982141

RESUMO

Adipose-tissue derived stromal cells (ASCs) are currently considered as a full value alternative source of bone marrow MSCs for prevention of graft-versus-host disease (GVHD) after hematopoietic stem cell transplantation due to their immunosuppressive potential. Besides, ASCs are known to support ex vivo expansion of hematopoietic stem and progenitor cells (HSPCs). Ex vivo expansion enables to amplify significantly the number of HSPCs of different commitment. Mononuclear cells (MNCs) from cord blood (cb) contain HSPCs and are easily assessed. The rarity of those HSPCs is a serious limitation of its application in cell therapy. Here we expanded cbMNCs in stroma-dependent setting to generate heterocellular associates consisting of ASCs and undifferentiated and low committed hematopoietic cbHSPCs. A part of cbHSPCs in associates demonstrated a primitive phenotype confirmed by formation of "cobblestone areas". ASCs associated with cbHSPCs demonstrated up-regulation of immunosuppressive indoleamine 2,3-dioxygenase (IDO), leukemia inhibitory factor (LIF), cyclooxygenase-2 (PTGS2) genes. ASC-cbHSPCs as well as ASCs provoked the suppression of HLA-DR activation and apoptosis of mitogen-stimulated T cells. VEGF transcription and secretion were elevated providing stimulation of blood vessel formation in ovo. Thus, ASCs retain immunosuppressive and proangiogenic capacities evidencing "third party" potential along with the effective support of ex vivo expansion of cbHSPCs. Above functions expand the relevance of ASCs for needs of regenerative medicine.


Assuntos
Tecido Adiposo/metabolismo , Técnicas de Cocultura/métodos , Sangue Fetal/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Células Estromais/metabolismo , Células Cultivadas , Sangue Fetal/citologia , Células-Tronco Hematopoéticas/citologia , Humanos , Células Estromais/citologia
16.
Arch Orthop Trauma Surg ; 140(4): 503-509, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31980879

RESUMO

INTRODUCTION: This study aimed to investigate the clinical outcomes after human umbilical cord blood-derived mesenchymal stem cell (hUCB-MSC) implantation for medial compartment (MC) osteoarthritis of the knee. MATERIALS AND METHODS: Inclusion criteria were patients older than 60 years, with a kissing lesion of the MC, a full-thickness chondral defect ≥ 4 cm2 of the medial femoral condyle (MFC), and a varus deformity ≥ 3° on a long cassette scanogram. The mean age was 64.9 ± 4.4 years and the mean chondral defect of the MFC was 7.2 ± 1.9 cm2. A mixture of sodium hyaluronate and hUCB-MSC was implanted into the chondral defect and a high tibial osteotomy was performed in all patients. International Knee Documentation Committee (IKDC), visual analog scale (VAS), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were evaluated preoperatively and 1 year and 2 years postoperatively. Cartilage regeneration was evaluated in 14 (56%) patients by second-look arthroscopy at 1 year postoperatively. RESULTS: Twenty-five patients underwent hUBC-MSC implantation. IKDC, VAS, and WOMAC scores at 1 year and 2 years improved significantly compared to preoperative scores. These scores at 1 year and 2 years were not significantly different between the body mass index (BMI) < 25 group and BMI ≥ 25 group. However, the < 65-year-old group showed superior IKDC scores at 1 year and 2 years and VAS score at 2 years than the ≥ 65-year-old group. Younger age and larger size of the chondral defect were associated with a significantly greater improvement in IKDC, VAS and WOMAC scores at 2 years. Second-look arthroscopy demonstrated International Cartilage Repair Society-Cartilage Repair Assessment grade I in six (42.9%) patients and grade II in eight (57.1%). CONCLUSIONS: hUCB-MSC implantation regenerated cartilage satisfactorily and showed satisfactory clinical outcomes in patients older than 60 years who had MC osteoarthritis.


Assuntos
Sangue Fetal/citologia , Transplante de Células-Tronco Mesenquimais , Osteoartrite do Joelho/cirurgia , Idoso , Artroscopia , Cartilagem Articular/cirurgia , Humanos , Articulação do Joelho/cirurgia , Pessoa de Meia-Idade , Cirurgia de Second-Look
17.
Lipids ; 55(1): 53-62, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31943229

RESUMO

The interest in the amount of polyunsaturated fatty acids (PUFA) in the umbilical cord blood (UCB) is increasing, but the stability of erythrocyte PUFA in these samples during storage and washing of the erythrocytes has not been directly evaluated. The purpose of this study was to analyze the effect of the lapse of time on the fatty acid (FA) content from UCB sample collection and maintained at 4 °C (0-12 h) until erythrocyte separation and washing. Palmitic acid (16:0), stearic acid (18:0), 18:1n-7/n-9, linoleic acid (18:2n-6), arachidonic acid (20:4n-6), 22:4n-6, eicosapentaenoic acid (20:5n-3), docosapentaenoic acid (22:5n-3), and docosahexaenoic acid (22:6n-3) together accounted for 87% of the FA profile in the umbilical vein erythrocytes. No difference was observed in the concentration of any of the FA studied, nor in the sum of saturated fatty acids (SFA), PUFA, or LC-PUFA in umbilical erythrocytes obtained at delivery and stored up to 12 h before the separation of erythrocytes. However, if a washing step was included in the processing of the erythrocytes, a decrease in the concentration of 16:0, 18:0, 18:3n-3, 20:4n-6, 22:4n-6, total SFA, PUFA, LC-PUFA, and n-6 LC-PUFA was evidenced, compared to unwashed erythrocytes. The FA concentration in umbilical cord erythrocytes did not change between samples stored from 0 to 12 h until erythrocyte separation. Erythrocyte washing before storage decreased the concentration of significant individual and total SFA, PUFA, and LC-PUFA. These results should be considered when planning the collection of UCB samples for the study of fatty acid concentration due to the nonscheduled timing of deliveries.


Assuntos
Eritrócitos/química , Ácidos Graxos/sangue , Sangue Fetal/citologia , Ácido Araquidônico/sangue , Ácidos Docosa-Hexaenoicos/sangue , Ácido Eicosapentaenoico/sangue , Ácidos Graxos/isolamento & purificação , Ácidos Graxos Insaturados/sangue , Feminino , Sangue Fetal/química , Idade Gestacional , Humanos , Ácido Linoleico/sangue , Ácido Palmítico/sangue , Gravidez , Ácidos Esteáricos/sangue
18.
Sci Rep ; 10(1): 1190, 2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-31988329

RESUMO

Pluripotent stem cell (PSC)-derived insulin-producing cells are a promising cell source for diabetes cellular therapy. However, the efficiency of the multi-step process required to differentiate PSCs towards pancreatic beta cells is variable between cell lines, batches and even within cultures. In adherent pancreatic differentiation protocols, we observed spontaneous local clustering of cells expressing elevated nuclear expression of pancreatic endocrine transcription factors, PDX1 and NKX6.1. Since aggregation has previously been shown to promote downstream differentiation, this local clustering may contribute to the variability in differentiation efficiencies observed within and between cultures. We therefore hypothesized that controlling and directing the spontaneous clustering process would lead to more efficient and consistent induction of pancreatic endocrine fate. Micropatterning cells in adherent microwells prompted clustering, local cell density increases, and increased nuclear accumulation of PDX1 and NKX6.1. Improved differentiation profiles were associated with distinct filamentous actin architectures, suggesting a previously overlooked role for cell-driven morphogenetic changes in supporting pancreatic differentiation. This work demonstrates that confined differentiation in cell-adhesive micropatterns may provide a facile, scalable, and more reproducible manufacturing route to drive morphogenesis and produce well-differentiated pancreatic cell clusters.


Assuntos
Diferenciação Celular/fisiologia , Sangue Fetal/citologia , Proteínas de Homeodomínio/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Células Secretoras de Insulina/metabolismo , Transativadores/metabolismo , Citoesqueleto de Actina/metabolismo , Adulto , Adesão Celular , Células Cultivadas , Diabetes Mellitus Tipo 1/terapia , Humanos , Transplante das Ilhotas Pancreáticas , Fenótipo , Reação em Cadeia da Polimerase em Tempo Real
19.
Artigo em Inglês | MEDLINE | ID: mdl-31926297

RESUMO

Maternal smoking-induced congenital heart and microvascular defects are closely associated with the impaired functioning of the in-utero feto-placental circulation system. Current groundbreaking facts revealed intimate crosstalk between circulating red blood cells (RBCs) and the vascular endothelium. Thus, RBCs have become the protagonists under varied pathological and adverse pro-oxidative cellular stress conditions. We isolated and screened fetal RBCs from the arterial cord blood of neonates, born to non-smoking (RBC-NS) and smoking mothers (RBC-S), assuming that parameters of fetal RBCs are blueprints of conditions experienced in-utero. Using atomic force microscopy and mass spectrometry-based shotgun lipidomics in the RBC-S population we revealed induced membrane stiffness, loss in intrinsic plastic activities and several abnormalities in their membrane-lipid composition, that could consequently result in perturbed hemodynamic flow movements. Altogether, these features are indicative of the outcome of neonatal microvascular complications and suggest unavailability for the potential rescue mechanism in cases of vascular endothelium impairment due to altered membrane integrity and rheological properties.


Assuntos
Eritrócitos/patologia , Sangue Fetal/citologia , Efeitos Tardios da Exposição Pré-Natal/etiologia , Poluição por Fumaça de Tabaco/efeitos adversos , Adulto , Fenômenos Biomecânicos , Membrana Eritrocítica/química , Membrana Eritrocítica/patologia , Eritrócitos/química , Feminino , Hemodinâmica , Humanos , Recém-Nascido , Peroxidação de Lipídeos , Fluidez de Membrana , Lipídeos de Membrana/análise , Gravidez , Efeitos Tardios da Exposição Pré-Natal/patologia , Adulto Jovem
20.
J Cell Physiol ; 235(2): 706-717, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31254289

RESUMO

Mesenchymal stem cells (MSCs) have been revealed to hold great potential for the development of new treatment approaches for various diseases. However, the clinical use of these cells is limited due to their tumorigenic effects. The therapeutic benefits of MSCs are largely dependent on paracrine factors including extracellular vesicles (EVs). EVs are nano-sized bilayer membrane structures containing lipids, microRNAs and proteins which play key roles in cell-to-cell communications. Because of their lower immunogenicity, tumorigenicity, and easier management, EVs have emerged as a new promising alternative to whole-cell therapy. Therefore, this paper reviews current preclinical studies on the use of EVs derived from human umbilical cord MSCs (hucMSCs) as a therapeutic approach in treatment of several diseases including neurological, cardiovascular, liver, kidney, and bone diseases as well as the cutaneous wound, inflammatory bowel disease, cancers, infertility, and other disorders.


Assuntos
Exossomos/metabolismo , Exossomos/transplante , Sangue Fetal/citologia , Células-Tronco Mesenquimais/metabolismo , Animais , Linhagem Celular Tumoral , Terapia Baseada em Transplante de Células e Tecidos/efeitos adversos , Micropartículas Derivadas de Células/metabolismo , Micropartículas Derivadas de Células/transplante , Modelos Animais de Doenças , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/transplante , Humanos , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Camundongos , Ratos , Cordão Umbilical/citologia
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