14.06.2018

Publication in ALTEX

Completely defined co-culture of adipogenic differentiated adipose-derived stem cells and microvascular endothelial cells

A.-C. Volz, L. Hack, F. Atzinger and P. J. Kluger

Abstract

Vascularized adipose tissue models are highly demanded as alternative to existing animal models to elucidate the mechanisms of widespread diseases, screen for new drugs or asses corresponding safety levels. Standardly used animal-derived sera therein, are associated to ethical concerns, the risk of contaminations and many uncertainties in their composition and impact on cells. Therefore, their use should be completely omitted. In this study, we developed a serum-free, defined co-culture medium and implemented it to set up an adipocyte/endothelial cell (EC) co-culture model.

Human adipose-derived stem cells were differentiated under defined conditions (diffASCs) and, like human microvascular ECs (mvECs), cultured in a developed defined co-culture medium in mono-, indirect or direct co-culture for 14 days. The developed defined co-culture medium was superior to compared mono-culture media and facilitated the functional maintenance and maturation of diffASCs including perilipin A expression, lipid accumulation and also glycerol and leptin release. The medium equally allowed mvEC maintenance, confirmed by the expression of cluster of differentiation 31 (CD31) and von Willebrand factor (vWF) and acetylated low-density lipoprotein (acLDL) uptake. Thereby mvECs showed a strong dependency on EC-specific factors. Additionally, the development of vascular structures by mvECs was facilitated when directly co-cultured with diffASCs.

The completely defined co-culture system allows for the serum-free setup of adipocyte/EC co-cultures and thereby represents a valuable and ethically acceptable tool for the setup of vascularized adipose tissue models.

More information.

DOI:10.14573/altex.1802191