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 for 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 10 healthy human donors and cultured under four different medium conditions (SCP, SPPP, SPPP + 1 nM basic fibroblast growth factor [bFGF], and 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 with 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.