College of Life Science, Sichuan University, Chengdu, China; National Engineering Laboratory for Oral Regenerative Medicine, Sichuan University, Chengdu, China; State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, China.
Artif Organs. 2014 Feb;38(2):E33-45. doi: 10.1111/aor.12162. Epub 2013 Sep 11.
Adipogenic differentiation of adipose-derived stem cells (ASCs) is known to be affected by many promoting and inhibiting factors, and correlated with surrounding cells and extracellular matrix. However, few studies have evaluated the effect of secreted biological molecules from adipose tissue explants on the growth of ASCs. In this study, ASCs were isolated and the secretory factors from adipose tissue explants (SFAE) were prepared from adipose tissue explants. The multilineage differentiation potential of ASCs was determined using different inductive media. The influence of SFAE on the colony formation and proliferation of ASCs was determined using colony-forming efficiency assay and Cell Counting Kit-8 assay, respectively. Real-time polymerase chain reaction and Western blot were performed to analyze the beneficial effect of SFAE on the adipogenesis and angiogenesis of ASCs. Utilizing gelatin scaffold, the influence of SFAE on ASCs was investigated in vivo. Subsequently, cell/gelatin constructs were cultured in a subcutaneous pocket on a rat dorsum for 2 and 9 weeks, and the resultant samples were histologically evaluated. Results showed that ASCs derived from adipose explants can differentiate along multiple lineages in vitro. Utilizing SFAE, the proliferation and colony-forming efficiency of ASCs were inhibited, while the expression of adipogenesis markers such as C/EBPβ, PPARγ2, and LPL, as well as angiogenesis factor VEGF-A were promoted. Moreover, the beneficial effect of SFAE on adipogenesis was revealed in vivo. In conclusion, our results suggested that SFAE has beneficial influence on adipogenesis and angiogenesis both in vitro and in vivo.
脂肪来源干细胞（ASCs）的成脂分化受许多促进和抑制因素的影响，并与周围细胞和细胞外基质相关。然而，很少有研究评估脂肪组织外植体分泌的生物分子对 ASCs 生长的影响。本研究从脂肪组织外植体中分离出 ASCs 并制备了脂肪组织外植体分泌因子（SFAE）。使用不同的诱导培养基确定 ASCs 的多能分化潜能。通过集落形成效率测定和细胞计数试剂盒-8 测定分别测定 SFAE 对 ASCs 集落形成和增殖的影响。实时聚合酶链反应和 Western blot 用于分析 SFAE 对 ASCs 成脂和成血管的有益作用。利用明胶支架，研究了 SFAE 对 ASCs 的体内影响。随后，将细胞/明胶构建体在大鼠背部的皮下口袋中培养 2 周和 9 周，并对所得样品进行组织学评估。结果表明，源自脂肪外植体的 ASCs 可以在体外沿着多个谱系分化。利用 SFAE，ASCs 的增殖和集落形成效率受到抑制，而成脂标志物如 C/EBPβ、PPARγ2 和 LPL 以及血管生成因子 VEGF-A 的表达则得到促进。此外，SFAE 在体内对成脂作用也具有有益效果。总之，我们的结果表明，SFAE 对体外和体内的成脂和成血管均具有有益的影响。
