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1.
Mol Reprod Dev ; 84(3): 229-245, 2017 03.
Artigo em Inglês | MEDLINE | ID: mdl-28044390

RESUMO

Derivation and stable maintenance of porcine induced pluripotent stem cells (piPSCs) is challenging. We herein systematically analyzed two piPSC lines, derived by lentiviral transduction and cultured under either leukemia inhibitory factor (LIF) or fibroblast growth factor (FGF) conditions, to shed more light on the underlying biological mechanisms of porcine pluripotency. LIF-derived piPSCs were more successful than their FGF-derived counterparts in the generation of in vitro chimeras and in teratoma formation. When LIF piPSCs chimeras were transferred into surrogate sows and allowed to develop, only their prescence within the embryonic membranes could be detected. Whole-transcriptome analysis of the piPSCs and porcine neonatal fibroblasts showed that they clustered together, but apart from the two pluripotent cell populations of early porcine embryos, indicating incomplete reprogramming. Indeed, bioinformatic analysis of the pluripotency-related gene network of the LIF- versus FGF-derived piPSCs revealed that ZFP42 (REX1) expression was absent in both piPSC-like cells, whereas it was expressed in the porcine inner cell mass at Day 7/8. A second striking difference was the expression of ATOH1 in piPSC-like cells, which was absent in the inner cell mass. Moreover, our gene expression analyses plus correlation analyses of known pluripotency genes identified unique relationships between pluripotency genes in the inner cell mass, which are to some extent, in the piPSC-like cells. This deficiency in downstream gene activation and divergent gene expression may be underlie the inability to derive germ line-transmitting piPSCs, and provides unique insight into which genes are necessary to achieve fully reprogrammed piPSCs. 84: 229-245, 2017. © 2016 Wiley Periodicals, Inc.


Assuntos
Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Fatores de Crescimento de Fibroblastos/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Células-Tronco Pluripotentes Induzidas/metabolismo , Fatores de Transcrição Kruppel-Like/metabolismo , Fator Inibidor de Leucemia/farmacologia , Animais , Células-Tronco Pluripotentes Induzidas/citologia , Suínos
2.
Physiol Genomics ; 48(1): 50-61, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26534934

RESUMO

A transcriptomic analysis of cultured human uterine smooth muscle cells (hUtSMCs) was performed to examine gene expression profiles in smooth muscle in an environment containing the two major steroid hormones that regulate the human myometrium in physiological states associated with estrous, pregnancy, labor, and pathophysiological states such as leiomyoma and endometrial cancer. hUtSMCs were treated with progesterone (P4) and 17ß-estradiol (E2) individually and in combination, in the presence and absence of RU486 (mifepristone). Transcription of many genes was modulated in the presence of P4 or E2 alone, but almost six times more genes were transcriptionally modulated in the presence of the P4/E2 hormone combination. In total 796 annotated genes were significantly differentially expressed in the presence of both P4 and E2 relative to their expression in untreated cells. Functional withdrawal of P4 by addition of RU486 effectively reversed almost all transcriptional changes caused by P4/E2 treatment. Gene ontology analysis of differentially expressed genes revealed a strong association between P4/E2 treatment and downregulated expression of genes involved in cell communication, signal transduction, channel activity, inflammatory response, and differentiation. Upregulated processes included cell survival, gene transcription, steroid hormone biosynthesis, muscle development, insulin receptor signaling, and cell growth.


Assuntos
Estradiol/farmacologia , Miócitos de Músculo Liso/metabolismo , Miométrio/citologia , Progesterona/farmacologia , Transcriptoma/efeitos dos fármacos , Sítios de Ligação , Células Cultivadas , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Ontologia Genética , Humanos , Mifepristona/farmacologia , Miócitos de Músculo Liso/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Fatores de Transcrição/metabolismo , Transcriptoma/genética
3.
BMC Genomics ; 16: 277, 2015 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-25888366

RESUMO

BACKGROUND: Genes and signalling pathways involved in pluripotency have been studied extensively in mouse and human pre-implantation embryos and embryonic stem (ES) cells. The unsuccessful attempts to generate ES cell lines from other species including cattle suggests that other genes and pathways are involved in maintaining pluripotency in these species. To investigate which genes are involved in bovine pluripotency, expression profiles were generated from morula, blastocyst, trophectoderm and inner cell mass (ICM) samples using microarray analysis. As MAPK inhibition can increase the NANOG/GATA6 ratio in the inner cell mass, additionally blastocysts were cultured in the presence of a MAPK inhibitor and changes in gene expression in the inner cell mass were analysed. RESULTS: Between morula and blastocyst 3,774 genes were differentially expressed and the largest differences were found in blastocyst up-regulated genes. Gene ontology (GO) analysis shows lipid metabolic process as the term most enriched with genes expressed at higher levels in blastocysts. Genes with higher expression levels in morulae were enriched in the RNA processing GO term. Of the 497 differentially expressed genes comparing ICM and TE, the expression of NANOG, SOX2 and POU5F1 was increased in the ICM confirming their evolutionary preserved role in pluripotency. Several genes implicated to be involved in differentiation or fate determination were also expressed at higher levels in the ICM. Genes expressed at higher levels in the ICM were enriched in the RNA splicing and regulation of gene expression GO term. Although NANOG expression was elevated upon MAPK inhibition, SOX2 and POU5F1 expression showed little increase. Expression of other genes in the MAPK pathway including DUSP4 and SPRY4, or influenced by MAPK inhibition such as IFNT, was down-regulated. CONCLUSION: The data obtained from the microarray studies provide further insight in gene expression during bovine embryonic development. They show an expression profile in pluripotent cells that indicates a pluripotent, epiblast-like state. The inability to culture ICM cells as stem cells in the presence of an inhibitor of MAPK activity together with the reported data indicates that MAPK inhibition alone is not sufficient to maintain a pluripotent character in bovine cells.


Assuntos
Massa Celular Interna do Blastocisto/metabolismo , Bovinos/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Mórula/metabolismo , RNA Mensageiro/metabolismo , Animais , Benzamidas/farmacologia , Massa Celular Interna do Blastocisto/efeitos dos fármacos , Bovinos/genética , Bovinos/metabolismo , Células Cultivadas , Difenilamina/análogos & derivados , Difenilamina/farmacologia , Técnicas de Cultura Embrionária , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Mórula/efeitos dos fármacos , Células-Tronco Pluripotentes/metabolismo , Inibidores de Proteínas Quinases/farmacologia
4.
Cancer Immunol Immunother ; 62(1): 51-63, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22736254

RESUMO

T cells may have a role in sustaining the leukemic clone in chronic lymphocytic leukemia (CLL). In this study, we have examined the ability of T cells from CLL patients to support the survival of the leukemic B cells in vitro. Additionally, we compared global gene expression of T cells from indolent CLL patients with healthy individuals and multiple myeloma (MM) patients. Apoptosis of purified leukemic B cells was inhibited in vitro when co-cultured with increasing numbers of autologous T cells (p < 0.01) but not autologous B and T cells of normal donors. The anti-apoptotic effect exceeded that of the anti-apoptotic cytokine IL-4 (p = 0.002) and was greater with CD8+ cells (p = 0.02) than with CD4+ cells (p = 0.05). The effect was depended mainly on cell-cell contact although a significant effect was also observed in transwell experiments (p = 0.05). About 356 genes involved in different cellular pathways were deregulated in T cells of CLL patients compared to healthy individuals and MM patients. The results of gene expression profiling were verified for 6 genes (CCL4, CCL5 (RANTES), XCL1, XCL2, KLF6, and TRAF1) using qRT-PCR and immunoblotting. Our results demonstrate that CLL-derived T cells can prevent apoptosis of leukemic B cells and have altered expression of genes that may facilitate the survival of the leukemic clone.


Assuntos
Apoptose/imunologia , Linfócitos B/imunologia , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Linfocítica Crônica de Células B/patologia , Linfócitos T/imunologia , Transcriptoma , Idoso , Idoso de 80 Anos ou mais , Técnicas de Cocultura , Feminino , Citometria de Fluxo , Humanos , Immunoblotting , Leucemia Linfocítica Crônica de Células B/genética , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
5.
Inhal Toxicol ; 25(11): 640-50, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24044680

RESUMO

Hypersensitivity pneumonitis (HP) represents the immunologically mediated lung disease induced by repeated inhalations of a wide variety of certain finely dispersed organic antigens. In susceptible subjects, these inhalations provoke a hypersensitivity reaction characterized by intense inflammation of the terminal bronchioles, the interstitium and the alveolar tree. The inflammation often organizes into granulomas and may progress to pulmonary fibrosis. Our previous work indicated that cell extract of gram-negative bacteria Pantoea agglomerans (SE-PA) causes, in young C57BL/6J mice, pulmonary changes that are very similar to the clinical manifestations of HP in men. The purpose of presented studies was to describe the response of mice immune system while exposed to SE-PA. Particular attention was paid to examine the age influence on SE-PA induced inflammation and fibrosis in lung tissue. We used 3- and 18-month-old C57BL/6J mice. Lung samples were collected from untreated mice and animals exposed to harmful agent for 7 and 28 days. HP development was monitored by histological and biochemical evaluation. Using ELISA tests, we examined concentration of pro- and anti-inflammatory cytokines in lung homogenates. Our study demonstrated again that SE-PA provokes in mice changes typical for the clinical picture of HP, and that successive stages of disease (acute, subacute and chronic) might be obtained by modulation of time exposure. Furthermore, we found that animals' age at the time of sensitization influences the nature of observed changes (cytokine expression pattern) and the final outcome (reaction intensity and scale of fibrosis).


Assuntos
Envelhecimento/imunologia , Alveolite Alérgica Extrínseca/imunologia , Misturas Complexas/toxicidade , Pantoea , Alveolite Alérgica Extrínseca/etiologia , Alveolite Alérgica Extrínseca/patologia , Animais , Citocinas/imunologia , Feminino , Hidroxiprolina/imunologia , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL
6.
Eur J Orthop Surg Traumatol ; 23(2): 169-76, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23412448

RESUMO

Regenerative medicine is a promising approach for addressing musculoskeletal disorders. Successful implementation of regenerative therapies is based upon existence of reliable, easy accessible cell sources. Mesenchymal tissues removed during total knee replacement (TKR) were investigated as a potential autologous stem cell source. Materials and methods Samples were collected from patients undergoing primary TKR mononuclear cells from adipose and synovial tissue; subchondral trabecular bone and osteoarthritic cartilage were isolated and assessed in terms of mesenchymal stem cells (MSC) content. Results MSCs obtained from all the investigated tissue types and from all donors showed proliferative, differentiation and surface markers characteristic of stemness. Important number of MSCs could be obtained in the first passage (P0). Mesenchymal tissues removed during TJR can qualitatively and quantitatively function as autologous MSC sources to be considered for regenerative therapies.


Assuntos
Articulação do Joelho/citologia , Células-Tronco Mesenquimais/metabolismo , Osteoartrite do Joelho/metabolismo , Tecido Adiposo/citologia , Idoso , Idoso de 80 Anos ou mais , Artroplastia do Joelho , Cartilagem Articular/citologia , Contagem de Células , Células Cultivadas , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/cirurgia , Projetos Piloto , Membrana Sinovial/citologia
7.
J Gene Med ; 12(2): 207-18, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-20082426

RESUMO

BACKGROUND: Mesenchymal stem cells (MSC) are widely regarded as a promising tool for cellular therapy applications, and genetic modification by safe, liposome-based vectors may enhance their therapeutic potential. METHODS: The present study describes the use of a cationic lipid vector (Lipofectamine 2000) to deliver genes to MSC isolated from a number of species in vitro and determined the characteristics of this vector system in terms of dose, toxicity and the time course of expression. In addition, the optimal use of a nuclear localization sequence (NLS) to enhance gene expression was explored. RESULTS: Lipofection of human MSC did not adversely affect their ability to differentiate into osteogenic- and adipogenic lineages. Although human and rat MSC were found to take up lipoplexes with relative efficiency, lower levels of gene expression were detected in rabbit MSC, demonstrating a crucial effect of species. Peptides containing reiterated motifs of NLS were found to significantly improve on the level of transgene expression. Optimal gene delivery was observed when a three-fold reiterated NLS sequence was included in the liposome formulation. CONCLUSIONS: Thus, nonviral gene delivery to MSC is feasible with efficiency being species dependent and can be enhanced by use of a three-fold reiterated NLS.


Assuntos
Regulação da Expressão Gênica , Células-Tronco Mesenquimais/metabolismo , Sinais de Localização Nuclear/metabolismo , Peptídeos/metabolismo , Transfecção/métodos , Sequência de Aminoácidos , Animais , Biomarcadores , Diferenciação Celular , Membrana Celular/metabolismo , Citometria de Fluxo , Proteínas de Fluorescência Verde/metabolismo , Humanos , Lipossomos , Luciferases/metabolismo , Células-Tronco Mesenquimais/citologia , Dados de Sequência Molecular , Sinais de Localização Nuclear/química , Peptídeos/química , Coelhos , Ratos
8.
Acta Neuropathol ; 119(5): 601-15, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-19967542

RESUMO

The blood brain barrier (BBB) is formed by capillary endothelial cells with inter-endothelial cell tight junctions and other cells such as pericytes and astrocytes present. Previous studies have shown a role for tight junction abnormalities in BBB leakage in multiple sclerosis (MS) brain. This marks a key stage in the development of inflammatory demyelination in MS. The aim of this study was to identify aberrantly expressed genes involved in BBB changes in MS lesions. A focused endothelial cell biology microarray, capable of detecting changes in expression of 113 endothelial cell-specific genes, was employed to analyse endothelial cell mRNA extracted from post-mortem control white matter, MS normal appearing white matter (NAWM), chronic active or inactive lesions by laser capture microdissection. Microarray analysis found 52 genes out of 113 analysed, predominantly in the activation functional group, to be differentially expressed in lesions compared to control or NAWM (p < 0.01). The majority of the differentially expressed genes were validated by quantitative real time PCR. In addition, the protein expression profiles of ICAM2, MMP2, and VEGFR1 were examined by immunofluorescent staining of selected tissue blocks. ICAM-2 was expressed at a higher level in chronic inactive lesions than control or NAWM, corresponding with the increased mRNA measured by microarray and real time PCR. The data shown, presenting a number of differentially expressed genes in the microvascular compartment of MS lesions, may shed light on the molecular mechanisms that are involved in the breakdown of the BBB. This moves us a step closer to the identification of potential therapeutic targets for repair of the compromised BBB.


Assuntos
Vasos Sanguíneos/metabolismo , Encéfalo/irrigação sanguínea , Células Endoteliais/metabolismo , Endotélio Vascular/metabolismo , Esclerose Múltipla/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Barreira Hematoencefálica/metabolismo , Encéfalo/metabolismo , Circulação Cerebrovascular/genética , Feminino , Expressão Gênica , Humanos , Imuno-Histoquímica , Masculino , Microdissecção/métodos , Microscopia Confocal , Pessoa de Meia-Idade , Esclerose Múltipla/metabolismo , Neurônios/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Junções Íntimas/genética , Junções Íntimas/metabolismo
9.
World J Stem Cells ; 11(1): 1-12, 2019 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-30705711

RESUMO

Cellular reprogramming and induced pluripotent stem cell (IPSC) technology demonstrated the plasticity of adult cell fate, opening a new era of cellular modelling and introducing a versatile therapeutic tool for regenerative medicine. While IPSCs are already involved in clinical trials for various regenerative purposes, critical questions concerning their medium- and long-term genetic and epigenetic stability still need to be answered. Pluripotent stem cells have been described in the last decades in various mammalian and human tissues (such as bone marrow, blood and adipose tissue). We briefly describe the characteristics of human-derived adult stem cells displaying in vitro and/or in vivo pluripotency while highlighting that the common denominators of their isolation or occurrence within tissue are represented by extreme cellular stress. Spontaneous cellular reprogramming as a survival mechanism favoured by senescence and cellular scarcity could represent an adaptative mechanism. Reprogrammed cells could initiate tissue regeneration or tumour formation dependent on the microenvironment characteristics. Systems biology approaches and lineage tracing within living tissues can be used to clarify the origin of adult pluripotent stem cells and their significance for regeneration and disease.

10.
World J Stem Cells ; 10(5): 43-56, 2018 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-29849930

RESUMO

The use of stem cells as carriers for therapeutic agents is an appealing modality for targeting tissues or organs of interest. Combined delivery of cells together with various information molecules as therapeutic agents has the potential to enhance, modulate or even initiate local or systemic repair processes, increasing stem cell efficiency for regenerative medicine applications. Stem-cell-mediated delivery of genes, proteins or small molecules takes advantage of the innate capability of stem cells to migrate and home to injury sites. As the native migratory properties are affected by in vitro expansion, the existent methods for enhancing stem cell targeting capabilities (modified culture methods, genetic modification, cell surface engineering) are described. The role of various nanoparticles in equipping stem cells with therapeutic small molecules is revised together with their class-specific advantages and shortcomings. Modalities to circumvent common challenges when designing a stem-cell-mediated targeted delivery system are described as well as future prospects in using this approach for regenerative medicine applications.

11.
Biol Open ; 7(8)2018 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-30026265

RESUMO

During early mammalian development, transient pools of pluripotent cells emerge that can be immortalised upon stem cell derivation. The pluripotent state, 'naïve' or 'primed', depends on the embryonic stage and derivation conditions used. Here we analyse the temporal gene expression patterns of mouse, cattle and porcine embryos at stages that harbour different types of pluripotent cells. We document conserved and divergent traits in gene expression, and identify predictor genes shared across the species that are associated with pluripotent states in vivo and in vitro Amongst these are the pluripotency-linked genes Klf4 and Lin28b The novel genes discovered include naïve- (Spic, Scpep1 and Gjb5) and primed-associated (Sema6a and Jakmip2) genes as well as naïve to primed transition genes (Dusp6 and Trip6). Both Gjb5 and Dusp6 play a role in pluripotency since their knockdown results in differentiation and downregulation of key pluripotency genes. Our interspecies comparison revealed new insights of pluripotency, pluripotent stem cell identity and a new molecular criterion for distinguishing between pluripotent states in various species, including human.

12.
Cell Cycle ; 16(11): 1070-1084, 2017 Jun 03.
Artigo em Inglês | MEDLINE | ID: mdl-28426281

RESUMO

Previous research has shown that a subpopulation of cells within cultured human dermal fibroblasts, termed multilineage-differentiating stress enduring (Muse) cells, are preferentially reprogrammed into induced pluripotent stem cells. However, controversy exists over whether these cells are the only cells capable of being reprogrammed from a heterogeneous population of fibroblasts. Similarly, there is little research to suggest such cells may exist in embryonic tissues or other species. To address if such a cell population exists in pigs, we investigated porcine embryonic fibroblast populations (pEFs) and identified heterogeneous expression of several key cell surface markers. Strikingly, we discovered a small population of stage-specific embryonic antigen 1 positive cells (SSEA-1+) in Danish Landrace and Göttingen minipig pEFs, which were absent in the Yucatan pEFs. Furthermore, reprogramming of SSEA-1+ sorted pEFs led to higher reprogramming efficiency. Subsequent transcriptome profiling of the SSEA-1+ vs. the SSEA-1neg cell fraction revealed highly comparable gene signatures. However several genes that were found to be upregulated in the SSEA-1+ cells were similarly expressed in mesenchymal stem cells (MSCs). We therefore termed these cells SSEA-1 Expressing Enhanced Reprogramming (SEER) cells. Interestingly, SEER cells were more effective at differentiating into osteocytes and chondrocytes in vitro. We conclude that SEER cells are more amenable for reprogramming and that the expression of mesenchymal stem cell genes is advantageous in the reprogramming process. This data provides evidence supporting the elite theory and helps to delineate which cell types and specific genes are important for reprogramming in the pig.


Assuntos
Reprogramação Celular , Embrião de Mamíferos/citologia , Fibroblastos/citologia , Fibroblastos/metabolismo , Antígenos Embrionários Estágio-Específicos/metabolismo , Animais , Biomarcadores/metabolismo , Cruzamento , Diferenciação Celular , Membrana Celular/metabolismo , Células Cultivadas , Endoglina/metabolismo , Perfilação da Expressão Gênica , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos , Sus scrofa
13.
Curr Stem Cell Res Ther ; 10(2): 90-8, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-24975620

RESUMO

Osteoarthritis (OA) is the most common degenerative joint disorder worldwide. OA represents an increasing threat to the quality of life of affected persons as well as for health resources expenditure. The incapability of cartilage to heal has been long time regarded as the major cause of progressive joint degeneration and functional impairment. Recent reports about the presence of progenitor cell populations within adult normal and OA cartilage invite to a reconsideration of the mechanisms involved in the onset and propagation of the disease as well as of the causes that are preventing the endogenous progenitors to recompose a functional extracellular matrix. The interplay between chronic joint inflammation, tissue functional and pathological load and the mechanosensitivity of progenitor cell populations are not yet fully understood. Elucidation of these complex correlations at molecular level could lead to identification of biomarkers for early detection, finding targets for the causal treatment of OA. The use of local progenitor populations in various cartilage regeneration strategies proves to be a fruitful avenue for research and clinical applications.


Assuntos
Células-Tronco Adultas/fisiologia , Osteoartrite/patologia , Células-Tronco Adultas/patologia , Animais , Fenômenos Biomecânicos , Cartilagem/patologia , Progressão da Doença , Terapia Genética , Humanos , Osteoartrite/metabolismo , Osteoartrite/terapia , Transdução de Sinais
14.
World J Stem Cells ; 7(1): 96-105, 2015 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-25621109

RESUMO

Musculoskeletal-related pain is one of the most disabling health conditions affecting more than one third of the adult population worldwide. Pain from various mechanisms and origins is currently underdiagnosed and undertreated. The complexity of molecular mechanisms correlating pain and the progression of musculoskeletal diseases is not yet fully understood. Molecular biomarkers for objective evaluation and treatment follow-up are needed as a step towards targeted treatment of pain as a symptom or as a disease. Stem cell therapy is already under investigation for the treatment of different types of musculoskeletal-related pain. Mesenchymal stem cell-based therapies are already being tested in various clinical trials that use musculoskeletal system-related pain as the primary or secondary endpoint. Genetically engineered stem cells, as well as induced pluripotent stem cells, offer promising novel perspectives for pain treatment. It is possible that a more focused approach and reassessment of therapeutic goals will contribute to the overall efficacy, as well as to the clinical acceptance of regenerative medicine therapies. This article briefly describes the principal types of musculoskeletal-related pain and reviews the stem cell-based therapies that have been specifically designed for its treatment.

15.
Stem Cell Res Ther ; 4(5): 118, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24079605

RESUMO

Despite multiple methods of treatment and a wealth of research in the field of regenerative medicine focusing on cartilage defects, the management of cartilage injuries remains a challenge. A recent study by Van Pham and colleagues proposes a method for preconditioning autologous adipose-derived stem cells. Their study offers evidence about the increased proliferative and chondrogenetic capabilities of platelet-rich plasma-treated adipose-derived stem cells and the increased efficiency of these in treating articular cartilage defects in mice. Even though the method needs further elaboration and the composition of the repair tissue requires investigation, the results are promising for the design of clinically acceptable cell therapies aimed at cartilage regeneration.


Assuntos
Tecido Adiposo/metabolismo , Engenharia Tecidual/métodos , Animais , Humanos
16.
Open Orthop J ; 7: 144-51, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23798982

RESUMO

Molecular medicine uses knowledge about cell structure and function for disease, diagnostics, stage characterisation and treatment. The advent of genomic technologies is considerably leading to developments in the field of molecular medicine. The accumulation of detailed information about gene expression, epigenetic variability, protein transcription and functional modulation is contributing to a new era in medicine. Rapid and early diagnostic procedures, molecular characterisation of degenerative and proliferative diseases and personalized therapies are predicted to lead to advancements in health prevention and treatment of disease. Diagnostic tools and therapies based on local and /or general modulation of cellular processes for traumatic or degenerative musculoskeletal conditions are becoming available. A logical consequence of the information derived from extensive data gathering, systems biology and systemic medicine has lead to significant improvements in understanding biological structure and function in a simultaneous bottom top and integrative, holistic manner. The description of disease mechanism at an intimate, subcellular level has a dual benefit. A thorough understanding of the crosstalk involved in molecular pathways both in the normal and the diseased state are expanding scientific knowledge and simultaneously are enabling design cell-targeted and individualized therapies. This paper presents a brief overview of current molecular based treatments available to the orthopedic surgeon and introduces the concept of systemic medicine from the perspective of musculoskeletal pathology.

17.
Curr Stem Cell Res Ther ; 7(5): 319-28, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22480416

RESUMO

INTRODUCTION: Osteoarthritis (OA) represents an increasing health issue worldwide. Regenerative medicine (RM) has raised the hope for introducing revolutionary therapies in clinical practice. Detection of autologus cell sources can improve accessibility to RM strategies. OBJECTIVES: To assess the presence and biological potential of mesehchymal stem cells in three tissues (subchondral bone, synovial layer, periarticular adipose tissue) in late stages osteoarthritic patients. MATERIAL AND METHODS: Samples were collected from subjects undergoing total knee replacement (TKR). MSCs were isolated and cultured in complete αMEM with ß FGF. Cell morphology and growth potential was assessed. Flow cytometry was used for detection of several relevant cell surface markers. Quantitative and qualitative assessment of differentiation potential towards three mesenchymal lineages (osteogenesis adipogenesis chondrogenesis) was performed. Time lapse life cell imaging of nondiferentiated cells over 24 hours period was used to determine cell kinetics. RESULTS: Mesenchymal cells derived from all donors and tissue types showed morphology, growth and surface cell markers associated with stemness. All cell types underwent differentiation toward three mesenchymal lineages with significant differences between tissues of origin, not between donors. Cell kinetics, as derived from life imaging records, was variable with tissue of origin, significant higher for adipose derived MSCS. CONCLUSION: Human late stage OA mesenchymal tissues, contain progenitors with proliferative and differentiation potential of MSCs. These populations can be used for research and autologus regenerative therapies. Further comparative studies with age matched non OA samples has the potential of contributing to deepening knowledge about disease occurrence and progression.


Assuntos
Células-Tronco Mesenquimais/fisiologia , Osteoartrite/patologia , Tecido Adiposo/patologia , Idoso , Diferenciação Celular , Processos de Crescimento Celular , Linhagem da Célula , Separação Celular , Feminino , Citometria de Fluxo , Humanos , Masculino , Células-Tronco Mesenquimais/citologia , Pessoa de Meia-Idade , Osteoartrite/metabolismo , Fenótipo , Medicina Regenerativa , Membrana Sinovial/patologia , Imagem com Lapso de Tempo
18.
J Biomol Screen ; 16(7): 744-54, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21593485

RESUMO

This study aims at generating immune chicken phage display libraries and single-chain antibodies (scFvs) specifically directed against cell surface markers of cultured peripheral blood mononuclear cells (PBMCs) that contain endothelial progenitor cells (EPCs). In contrast to previous approaches that use well-defined recombinant antigens attached to plastic surfaces that may alter the structure of the proteins, the authors describe a method that maintains the cell surface markers on live cells while providing the opportunity to rapidly screen entire libraries for antibodies that bind to unknown cell surface markers of progenitor/stem cells. Chickens immunized with live EPCs, consisting of a heterogeneous population of lymphocytes and monocytes, demonstrated a robust immune response. After three rounds of biopanning, the authors purified and characterized three unique scFvs called UG1-3. Codon-optimized recombinant UG1 (gUG-1) shows binding by flow cytometry to circulating CD14-positive cells in peripheral blood consistent with predominant expression of a target protein on monocyte subsets. The authors describe the successful use of immunization of chickens for the generation of scFvs against a heterogenous population of EPCs displaying unknown cell surface markers and demonstrate the strong potential of phage display technology in the development of reagents for the isolation and characterization of stem/progenitor cells.


Assuntos
Células Endoteliais/imunologia , Biblioteca de Peptídeos , Anticorpos de Cadeia Única/imunologia , Células-Tronco/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Superfície/imunologia , Antígenos de Superfície/metabolismo , Galinhas/imunologia , Células Endoteliais/metabolismo , Epitopos/imunologia , Citometria de Fluxo , Humanos , Imunização , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Dados de Sequência Molecular , Ligação Proteica/imunologia , Alinhamento de Sequência , Anticorpos de Cadeia Única/isolamento & purificação , Células-Tronco/metabolismo
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