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Isolation and Characterization of Multipotent and Pluripotent Stem Cells from Human Peripheral Blood

机译:人外周血多能和多能干细胞的分离与鉴定

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Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.
机译:干细胞通常根据其分离的发育阶段进行分类,尽管这已引起干细胞科学家的争论。常用方法将干细胞分为三类:胚胎干细胞(ESC),脐带干细胞(UCBSC)和成人干细胞(ASC),其中包括骨髓干细胞(BM),脂肪组织(FT),工程诱导多能性(IP)和外周血(PB)。根据定义,干细胞是能够自我更新和分化的假想祖细胞,假设“无限”(ab infinitum)成为更专门的细胞和组织。这项研究的主要目的是确定和表征人PB-SC中存在的干细胞的不同亚群,这可能代表了临床再生医学领域的一个有效机会。使用多学科方法对分离的单核细胞中的干细胞进行表征,该方法基于形态学,通过流式细胞术和荧光分析的干细胞标志物表达,RT-PCR以及自我更新或增殖并分化为专门细胞的能力。该方法用于鉴定造血,间充质,胚胎和神经干细胞标志物的表达。分离的贴壁细胞和悬浮单核细胞都能够通过在暴露于适当的诱导培养基时分别保持其增殖和分化为成骨细胞的能力,从而在体外培养过程中保持干细胞特性。

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