Department of Oral & Maxillofacial Surgery, Christian-Albrechts-University, Kiel, Germany.
Department of Oral & Maxillofacial Surgery, Christian-Albrechts-University, Kiel, Germany.
J Plast Reconstr Aesthet Surg. 2014 Oct;67(10):1427-35. doi: 10.1016/j.bjps.2014.05.042. Epub 2014 May 29.
Adipose-derived stromal cells (ASCs) are mostly isolated by enzymatic digestion, centrifugation and adherent growth resulting in a very heterogeneous cell population. Therefore, other cell types in the cell culture can comprise the differentiation and proliferation potential of the ASC population. Recent studies indicated that an antibody-aided isolation of distinct ASC subpopulations provides advantages over the conventional method of ASC isolation. The aim of this study was to investigate the adipogenic differentiation potential of CD29-, CD71-, CD73- and CD90-selected ASCs in vitro. The stromal vascular fraction (SVF) was obtained from rat adipose tissue by enzymatic digestion and centrifugation. Subsequently, CD29(+)-, CD71(+)-, CD73(+)- and CD90(+) cells were isolated by magnetic activated cell sorting (MACS), seeded into culture plates and differentiated into the adipogenic lineage. ASCs isolated by adherent growth only served as controls. Adipogenic differentiation was assessed by Oil Red O staining and quantification of the adiponectin and leptin concentrations in the cell culture supernatants. Statistical analysis was carried out using one-way analysis of variance (ANOVA) followed by the Scheffe's post hoc procedure. The results showed that different subpopulations with different adipogenic differentiation potentials can be isolated by the MACS procedure. The highest adipogenic differentiation potential was determined in the CD29-selected ASC population followed by the unsorted ASC population. The CD71-, CD73- and CD90-selected cells exhibited significantly the lowest adipogenic differentiation potential. In conclusion, the CD29-selected ASCs and the unsorted ASCs exhibited a similar adipogenic differentiation potential. Therefore, we do not see a clear advantage in the application of an anti-CD29-based isolation of ASCs over the conventional technique using adherent growth. However, the research on isolation/purification methods of adipogenic ASCs should continue in order to make this stem cell source even more attractive for future adipose tissue engineering applications.
脂肪来源的基质细胞（ASCs）主要通过酶消化、离心和贴壁生长分离，从而产生非常异质的细胞群体。因此，细胞培养中的其他细胞类型可能包含 ASC 群体的分化和增殖潜力。最近的研究表明，与 ASC 分离的传统方法相比，抗体辅助分离特定的 ASC 亚群具有优势。本研究旨在研究体外 CD29-、CD71-、CD73- 和 CD90-选择的 ASC 的成脂分化潜力。基质血管部分（SVF）通过酶消化和离心从大鼠脂肪组织中获得。随后，通过磁性激活细胞分选（MACS）分离 CD29(+)、CD71(+)、CD73(+) 和 CD90(+)细胞，接种到培养板中并分化为成脂谱系。仅通过贴壁生长分离的 ASC 作为对照。通过油红 O 染色和细胞培养上清液中脂联素和瘦素浓度的定量评估成脂分化。使用单向方差分析（ANOVA）进行统计分析，然后进行 Scheffe 事后检验。结果表明，通过 MACS 程序可以分离具有不同成脂分化潜力的不同亚群。CD29 选择的 ASC 群体具有最高的成脂分化潜力，其次是未分选的 ASC 群体。CD71、CD73 和 CD90 选择的细胞表现出明显最低的成脂分化潜力。总之，CD29 选择的 ASC 和未分选的 ASC 表现出相似的成脂分化潜力。因此，我们在应用基于抗-CD29 的 ASC 分离方法方面没有看到明显的优势，而传统的使用贴壁生长的技术。然而，应该继续研究成脂 ASC 的分离/纯化方法，以使这种干细胞来源更具吸引力，用于未来的脂肪组织工程应用。
