RESUMO
Approximately 90 % of infants born before 28 full weeks(extremely-preterm-infants) receive erythrocyte transfusions in early life. Umbilical cord blood(UCB) has been investigated as an alternative source for erythrocyte transfusions to preterm neonates. This retrospective study aimed to compile/evaluate spectrum of bacteria groups/species intermittently detected in processed UCB at National-Swedish-Cord blood bank, (NS-CBB) during the years 2008-2020. Consecutive data from the years 2008-2020 were investigated. UCB from healthy newborns born after 37 full weeks of gestation was collected following clamping of cord (1 min) through cannulation of umbilical vein(vaginal-and C-section-deliveries). In total, 5194 cord blood units (UCBUs) that met NS-CBB-guidelines for total nucleated-cell-content(TNC) were manufactured from 8875 collections. Of 5194 UCBUs,77,6 % were from vaginal-and 22,4 % from C-section deliveries.Samples(10 mL) were collected from surplus eryhtrocyte fraction post-processing(n = 5194), transferred into BACT/ALERT® aerobic/anaerobic culture flasks and monitored 10 days using BACT/ALERT®-3D-Microbial-Detection-Systems. Positive samples were subcultured and typed for bacterial groups and/or species. Out of 5194 processed sampled UCB units,186 (3,6 %) were discarded due to positive sterility tests, 92 % were detected in samples from vaginal-deliveries and 8 % from C-section-deliveries. In all,16 different groups of bacteria and 27 species were identified. Common bacterial/groups and species were anaerobe gram-negative rods(n = 28),coagulase-negative-staphylococci(n = 21),gram-positive rods(n = 21),anaerobe-gram-positive cocci(n = 20) and viridans-streptococci(n = 13). Extracted from these results,in positive samples(n = 13) from C-section deliveries, bacteria were found:viridans-streptococci(n = 7),Aerococcus-urinae(n = 1), Staphylococcus lugdunensis(n = 1),other coagulase-negative staphylococci(n = 1) or a mix of aerobic/anaerobic bacteria(n = 3). Our results are in alignment with previously published contamination rates in processed UCBUs. Still, results point towards importance of strict microbial monitoring when manufacturing UCBUs to achieve patient-safe- products for stem-cell transplantation/transfusion.
Assuntos
Sangue Fetal , Humanos , Estudos Retrospectivos , Recém-Nascido , Feminino , Bactérias , MasculinoRESUMO
Low back pain is a major health issue and one main cause to this condition is believed to be intervertebral disc (IVD) degeneration. Stem cell therapy for degenerated discs using mesenchymal stromal cells (MSCs) has been suggested. The aim of the study was to investigate the presence and distribution pattern of autologous MSCs transplanted into degenerated IVDs in patients and explanted posttransplantation. IVD tissues from four patients (41, 45, 47, and 47 years of age) participating in a clinical feasibility study on MSC transplantation to degenerative discs were investigated. Three patients decided to undergo fusion surgery at time points 8 months and one patient at 28 months posttransplantation. Pretransplantation, MSCs from bone marrow aspirate were isolated by centrifugation in FICOLL® test tubes and cultured (passage 1). Before transplantation, MSCs were labeled with 1 mg/mL iron sucrose (Venofer®) and 1 × 106 MSCs were transplanted into degenerated IVDs. At the time point of surgery, IVD tissues were collected. IVD tissue samples were fixated, embedded in paraffin, and sections prepared. IVD samples were stained with Prussian Blue, by which iron deposits are visualized and examined (light microscopy). Immunohistochemistry (IHC), including SOX9 (sex determining region Y box 9), Coll2A1 (collagen 2A1), and cell viability (TUNEL) were performed. Cells positive for iron deposits were observed in IVD tissues (3/4 patients). The cells/iron deposits were observed in clusters and/or as solitary cells in regions in IVD tissue samples [regions of interest (ROIs)]. By IHC, SOX9- and Coll2A1-positive cells were detected in the same regions as the detected cells/iron deposits. A few nonviable cells were detected by TUNEL assay in ROIs. Results demonstrated that MSCs, labeled with iron sucrose, transplanted into degenerated IVDs were detectable 8 months posttransplantation. The detected cellular activity indicates that MSCs have differentiated into chondrocyte-like cells and that the injected MSCs and/or their progeny have survived since the cells were found in large cluster and as solitary cells which were distributed at different parts of the IVD.
Assuntos
Degeneração do Disco Intervertebral/terapia , Dor Lombar/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , Adulto , Movimento Celular , Células Cultivadas , Condrogênese , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Feminino , Humanos , Injeções Espinhais , Degeneração do Disco Intervertebral/patologia , Ferro/metabolismo , Dor Lombar/patologia , Masculino , Transplante de Células-Tronco Mesenquimais/efeitos adversos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Pessoa de Meia-Idade , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismoRESUMO
Physical exercise has several beneficial effects on the heart. In other tissues it has been shown to activate endogenous stem cells. There is however a lack of knowledge how exercise affects the distribution of progenitor cells as well as overall cell turnover within the heart. Therefore, proliferating cells were identified using BrdU DNA labeling in a rat exercise model. Slow cycling cells were identified by label retention. BrdU+ nuclei were counted in apex, ventricle and atrioventricular junction (AV junction), as well as in skin tissue where label retaining cells (LRC) have been described previously. After 13 weeks of chasing, the cells with the highest intensity were identified and considered as LRC. Heart tissue showed slower proliferation compared to skin. The highest number of BrdU+ cells was found in the AV junction. Here, a sub region in close proximity to the valvular insertion point was observed, where density of BrdU+ cells was high at all time points. Physical exercise increased proliferation in AV junction at the early stage. Furthermore, the sub region was found to harbor a significant higher number of LRC compared to other regions of the heart in the exercised animals. Progenitor markers MDR1 and Sca-1 were detected in the same area by immunohistochemistry. In conclusions, our data shows that physical exercise affects cell turnover and distribution of LRC in the heart. Furthermore, it reveals a region within the AV junction of the heart that shows features of a stem cell niche.
Assuntos
Ciclo Celular , Microambiente Celular , Miocárdio/metabolismo , Miócitos Cardíacos/metabolismo , Condicionamento Físico Animal , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Biomarcadores , Feminino , Imuno-Histoquímica , Ratos , Nicho de Células-Tronco , Células-Tronco/metabolismoRESUMO
At the present time, the normal cell proliferation rate and regeneration processes in the intervertebral disc (IVD) are not fully known. Historically, the IVD has been considered an organ with little or no regenerative capacity. However, several studies have identified the presence of cells expressing progenitor/stem cell markers in adult cartilage tissue and recent data suggest that adult mammalian IVDs have regenerative capacity, albeit slow. The aim of this review is to give an overview of the present knowledge regarding IVD development, regeneration and repair mechanisms in mammals, with a special focus on human discs. At a time when regenerative medicine is making progress and biological treatment options, such as stem cell therapy, are suggested for patients with degenerated discs causing chronic low back pain, basic knowledge about disc cells and their regenerative capacity form a useful basis for the exploration of new treatment options.
Assuntos
Disco Intervertebral/fisiologia , Regeneração/fisiologia , Animais , Humanos , Disco Intervertebral/crescimento & desenvolvimento , Disco Intervertebral/metabolismo , Mamíferos , Medicina RegenerativaRESUMO
STUDY DESIGN: Descriptive experimental study. OBJECTIVE: The aim of this study was to investigate the effect of exercise on cell proliferation in different areas of the intervertebral disc (IVD) and recruitment of cells possibly active in regeneration of normal rat lumbar IVDs. SUMMARY OF BACKGROUND DATA: Little is known about the effects of physical exercise on lumbar IVD tissue. Recently, stem cell niches in the perichondrium area of the IVD were identified and cells in these niches have been suggested to be involved in the normal regeneration of the IVD. METHODS: Thirty Sprague-Dawley rats were exposed to 5-bromo-2-deoxyuridine (BrdU) diluted in the drinking water during 14 days. Fifteen rats ran on a treadmill daily for 50 min/d, 5 d/wk (exercise group), and 15 nonexercised rats served as controls. Immunohistochemical analyses (anti-BrdU antibody) were performed at 9, 14, 28, 56, and 105 days after the start of the exercise protocol. BrdU positive cells were counted in the stem cell niche area, the peripheral region of epiphyseal cartilage area, and the annulus fibrous outer and inner areas. Data were analyzed by 2-way analysis of variance (significance level; P < 0.05). RESULTS: The BrdU positive cell numbers in the stem cell niche and annulus fibrous outer regions were increased in discs from the exercising group on days 14 (P < 0.01) and 105 (P < 0.05) and at day 14 (P < 0.01) in the peripheral epiphyseal cartilage region compared with controls. CONCLUSION: Physical exercise was shown to have positive effects on cell proliferation in IVDs, with involvement of various disc regions, indicating a differential response by disc tissue to exercise depending on anatomical location and tissue characteristics.
Assuntos
Proliferação de Células , Disco Intervertebral/citologia , Vértebras Lombares/citologia , Condicionamento Físico Animal/fisiologia , Animais , Bromodesoxiuridina/metabolismo , Bromodesoxiuridina/farmacologia , Contagem de Células , Feminino , Lâmina de Crescimento/citologia , Imuno-Histoquímica , Disco Intervertebral/efeitos dos fármacos , Disco Intervertebral/metabolismo , Ratos , Ratos Sprague-Dawley , Nicho de Células-Tronco/efeitos dos fármacos , Fatores de TempoRESUMO
STUDY DESIGN: Descriptive experimental study performed in rabbits of 2 age groups. OBJECTIVE: To study and investigate presence of prechondrocytic cells and cell migration routes (MR) in the intervertebral disc (IVD) region to gain knowledge about the normal IVD regeneration pattern. SUMMARY OF BACKGROUND DATA: Disc degeneration is thought to play a major role in patients with chronic lumbar pain. Regeneration processes and cell migration within the IVD have been sparsely described. Therefore, it is of interest to increase knowledge of these processes in order to understand pathological conditions of the IVD. METHODS: At the beginning of the experiment, 5-bromo-2-deoxyuridine (BrdU) in vivo labeling was performed in 2 groups of rabbits, 3 and 9 months old (total 27 rabbits). BrdU is incorporated into DNA during mitosis, and then it is gradually diluted with each cell division until it finally disappears. Incorporation of BrdU was then visualized by immunohistochemistry (IHC) at different time points providing cell division pattern and presence of slow-cycling cells in the IVD region. IVD tissue was investigated by IHC for growth and differentiation factor-5 (GDF5), SOX9 (chondrogenic lineage markers), SNAIL homolog 1 (SNAI1), SNAIL homolog 2 (SLUG) (migration markers), and ß1-INTEGRIN (cellular adhesion marker). In addition, GDF5, SOX9, and BMPRIB expression were investigated on genetic level. RESULTS: BrdU cells were observed in early time points in the IVD niche, adjacent to the epiphyseal plate, at later time points mainly in outer region of the annulus fibrosus for both age groups of rabbits, indicating a gradual migration of cells. The presence of SLUG, SNAI1, GDF5, SOX9, and ß1-INTEGRIN was found in same regions. CONCLUSION: The results suggest a cellular MR from the IVD stem cell niche toward the annulus fibrosus and the inner parts of the IVD. These findings may be of importance for understanding IVD regenerative mechanisms and for future development of biological treatment strategies.
Assuntos
Envelhecimento/fisiologia , Movimento Celular , Disco Intervertebral/crescimento & desenvolvimento , Regeneração , Nicho de Células-Tronco , Células-Tronco/fisiologia , Fatores Etários , Envelhecimento/genética , Envelhecimento/metabolismo , Animais , Biomarcadores/metabolismo , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/genética , Receptores de Proteínas Morfogenéticas Ósseas Tipo I/metabolismo , Diferenciação Celular , Linhagem da Célula , Proliferação de Células , Condrogênese , Replicação do DNA , Feminino , Regulação da Expressão Gênica , Fator 5 de Diferenciação de Crescimento/genética , Fator 5 de Diferenciação de Crescimento/metabolismo , Imuno-Histoquímica , Integrina beta1/metabolismo , Disco Intervertebral/metabolismo , Coelhos , Fatores de Transcrição SOX9/genética , Fatores de Transcrição SOX9/metabolismo , Fatores de Transcrição da Família Snail , Células-Tronco/metabolismo , Fatores de Transcrição/metabolismoRESUMO
A superficial lesion of the articular cartilage does not spontaneously self-repair and has been suggested to be partly due to lack of progenitor cells within the joint that can reach the site of injury. To study whether progenitor cells are present within the joint, 3-month-old New Zealand white rabbits were exposed to bromodeoxyuridine (BrdU) for 12 consecutive days and were then sacrificed 4, 6, 10, 14, 28 and 56 days after the first BrdU administration. Presence of BrdU and localization of progenitor markers were detected using immunohistochemistry. After 10 days of BrdU exposure, BrdU-positive cells, i.e. proliferating cells, were abundantly detected in the epiphyseal plate, the perichondrial groove of Ranvier, and in all zones of the articular cartilage. After a wash-out period, BrdU-positive cells were still present, i.e. those considered to be progenitor cells, in these regions of the knee except for the proliferative zone of the epiphyseal plate. Cells in the perichondrial groove of Ranvier were further positive for several markers associated with progenitor cells and stem cell niches, including Stro-1, Jagged1, and BMPr1a. Our results demonstrate that a small population of progenitor cells is present in the perichondrial groove of Ranvier as well as within the articular cartilage in the knee. The perichondrial groove of Ranvier also demonstrates the properties of a stem cell niche.