Human serum from platelet-poor plasma for the culture of primary human preadipocytes

¹ Department of Plastic Surgery and Hand Surgery, Burns Center, University Hospital, Aachen University of Technology, Aachen, Germany

^* To whom correspondence should be addressed. E-mail: E.Koellensperger{at}gmx.net.

	Abstract

In adipose tissue engineering, the use of human serum is essential to achieve the goal of an autologous system. Serum from conventional human plasma (SCP) contains platelet-derived growth factor (PDGF), a growth factor known to be both a potent inhibitor of adipose differentiation and also the most important stimulator of proliferation in human serum. Serum from platelet-poor plasma (SPPP) is considered to be PDGF-deprived, and should therefore inhibit the differentiation of preadipocytes to adipocytes to a lesser extent. Effective cultivation of preadipocytes with SPPP requires compensating the missing stimulatory PDGF effect on proliferation. However, the addition of other growth factors to the media needs to provide stimulation of proliferation without significant inhibition of differentiation. Primary human preadipocytes were isolated from adipose tissue samples of ten healthy human donors and cultured under four different media conditions [SCP, SPPP, SPPP + 1 nM basic fibroblast growth factor (bFGF), SPPP + 1 nM epidermal growth factor (EGF)] for five generations. Proliferation activity and differentiation capacity were assessed for each sample, generation and culture condition by calculating doubling time and measuring glycerol 3-phosphate-dehydrogenase (GPDH)-specific activity. The use of SPPP resulted in a marked rise in GPDH activity compared to the cells cultured with SCP. Supplementing SPPP with 1 nM bFGF or EGF increased proliferation activity significantly. SPPP can be considered superior to SCP for the culture of primary human preadipocytes in adipose tissue engineering in terms of proliferation activity and differentiation capacity.

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