Effective harvesting procedure for adipose tissue is demanded by the affordable Good Manufacturing Practice-Compliant Production of clinical-grade adipose tissue-derived stem cells (hADSCs). Enzymatic digestion using collagenase is the most reliable method of adipose tissue-derived stem cells (hADSCs) isolation, while the optimized loading volume ratios of digestion to container during the shaking process of adipose tissue and collagenase mixture are still lacking. This study was conducted to determine the optimized loading volume ratio (mixture to container) for enzymatic digestion of Stromal/Stem Cells from lipoaspirate. Lipoaspirates were obtained from twelve women immediately after liposuction. Then tissue from each patient was divided into four groups according to different loading volume ratios in 50 ml centrifugal tube: 0.2 group, 0.4 group, 0.6 group, 0.8 group. Stromal vascular fractions (SVF) were obtained from each group, then total cell counts, viability and viable cell count were performed. hADSCs were harvested at passage (P) 2, whose morphologies, immunophenotypes, proliferation, and tri-differentiation abilities were compared. 0.4 loading volume ratio provided the highest cell yield, favorable viability and viable cell yield. The proliferation and triple differentiation ability of hADSCs obtained by 0.4 group was not inferior to that of other groups. Therefore, 0.4 may be the optimal loading volume ratio for hADSCs isolation from lipoaspirate by enzymatic digestion in current setting.

中文翻译：

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不同消化负荷比的脂肪抽吸物中基质/干细胞的细胞产量和生物学特性。


临床级脂肪组织干细胞 (hADSC) 的生产符合良好生产规范，需要有效的脂肪组织采集程序。使用胶原酶进行酶消化是分离脂肪组织源性干细胞（hADSC）最可靠的方法，但在脂肪组织和胶原酶混合物的摇动过程中消化与容器的最佳装载体积比仍然缺乏。本研究的目的是确定从脂肪抽吸物中酶消化基质/干细胞的最佳装载体积比（混合物与容器）。抽脂术后立即从 12 名女性身上获取抽脂物。然后将每位患者的组织按照50 ml离心管中不同装载体积比例分为四组：0.2组、0.4组、0.6组、0.8组。从每组中获得基质血管组分（SVF），然后进行总细胞计数、活力和活细胞计数。在第(P)2代收获hADSC，比较其形态、免疫表型、增殖和三分化能力。 0.4的装载体积比提供了最高的细胞产量、良好的活力和活细胞产量。 0.4组获得的hADSCs的增殖能力和三向分化能力并不逊色于其他组。因此，在当前设置下，0.4 可能是通过酶消化从脂肪抽吸物中分离 hADSC 的最佳装载体积比。