Optimierung der Proliferation und Differenzierung humaner Präadipozyten durch den Einsatz von humanem Serum

摘要

Reconstruction of soft tissue defects is a challenge in plastic surgery and there is clinical need for adequate solutions. A promising solution is adipose tissue engineering with preadipocytes. These adipogenic precursor cells have the ability to proliferate and differentiate into adipose tissue after transplantation. This study aimed at developing an optimal protocol for proliferation and differentiation of preadipocytes. In this study, the yield of preadipocytes obtained from liposuction material was compared to that of excised adipose tissue. Furthermore, we compared the two media DMEM and OptiMEM and their effects on proliferation and adipogenic conversion. We also investigated the effects of FCS versus human serum on preadipocyte proliferation and differentiation. In addition we evaluate the advantages of fibronectin-coating concerning the yield of preadipocytes after isolation from adipose tissue as well as the effects of fibronectin on preadipocyte differentiation. Our results show clearly that the yield of preadipocytes isolated from liposuction material is higher than the yield from excised adipose tissue. Our findings show how the use of human serum for preadipocyte cultivation allows substantial differentiation of adipocyte precursors even after an extensive in vitro expansion. The results also show that fibronectin coating does not only strongly increase the yield of preadipocytes after isolation from adipose tissue but also significantly enhances differentiation of precursor cells to mature adipocytes. Our results smooth the way for autologous preadipocyte culturing and show that human serum for preadipocyte culturing opens new and promising perspectives for adipose tissue engineering by optimizing in vitro expansion in cell culture and inducing substantial differentiation.