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Lineage depletion of stromal vascular fractions isolated from human adipose tissue: a novel approach towards cell enrichment technology

机译：从人类脂肪组织中分离出的基质血管部分的谱系耗竭：一种新的细胞富集技术

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The therapeutic rationale for tissue repair and regeneration using stem cells is at its infancy and needs advancement in understanding the role of individual component’s innate capability. As stem cells of adipose tissue reside in a more heterogeneous population of stromal vascular fractions, cell separation or sorting becomes an eminent step towards revealing their unique properties. This study elucidates the comparative efficacy of lineage depleted adipose derived stromal vascular fraction (SVF) and their innate ability using magnetic activated cell sorter (MACS). To this end, isolated SVF from human adipose tissue was lineage depleted according to the manufacturer’s instructions using specific antibody cocktail through MACS. The enriched lineage negative (lin?) and lineage positive (lin+) cell fractions were cultured, phenotypically characterized for the panel of cell surface markers using flowcytometry and subjected to osteoblastic and adipogenic differentiation. The expression profile obtained for lin? cells was CD34?/CD45?/HLADR?/CD49d?/CD140b?/CD31?/CD90+/CD105+/CD73+/CD54+/CD166+/CD117? when compared to Lin+ cells expressing CD34+/CD45+/HLADR?/CD49d?/CD140b+/CD31?/CD90+/CD105+/CD73+/CD54+/CD166+/CD117+ (CD—cluster of differentiation). These results, thus, advances our understanding on the inherent property of the individual cell population. Furthermore, both the fractions exhibited mesodermal lineage differentiation capacity. To conclude, this research pursuit rationalized the regenerative therapeutic applicability of both lin? and lin+ cultures of human adipose tissue for disorders of mesodermal, haematological and vascular origin.
机译：使用干细胞进行组织修复和再生的治疗原理尚处于起步阶段，需要进一步了解单个组件的先天能力的作用。随着脂肪组织干细胞驻留在基质血管部分的异质性群体中，细胞分离或分选成为揭示其独特性质的重要步骤。这项研究阐明了沿谱耗竭的脂肪来源的基质血管部分（SVF）的比较功效及其使用磁性激活细胞分选仪（MACS）的先天能力。为此，根据制造商的说明，使用了通过MACS的特异性抗体混合物，从人类脂肪组织中分离出了SVF谱系。培养富集的谱系阴性（lin +）和谱系阳性（lin +）细胞级分，使用流式细胞仪对细胞表面标志物进行表型鉴定，并进行成骨和成脂分化。获得的表达谱为lin？细胞是CD34？/ CD45？/ HLADR？/ CD49d？/ CD140b？/ CD31？/ CD90 + / CD105 + / CD73 + / CD54 + / CD166 + / CD117？与表达CD34 + / CD45 + /HLADRα/CD49dβ/ CD140b + /CD31β/ CD90 + / CD105 + / CD73 + / CD54 + / CD166 + / CD117 +（CD-分化群）的Lin +细胞相比时。因此，这些结果使我们对单个细胞群的固有特性有了更深入的了解。此外，两个部分均显示出中胚层谱系分化能力。总而言之，这项研究追求合理化了两种林的再生治疗适用性。和人类脂肪组织的lin +培养物用于中胚层，血液学和血管源性疾病。

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