Clinic for Plastic and Reconstructive Surgery, Aesthetic and Preventive Medicine at Heidelberg University Hospital-ETHIANUM, Heidelberg, Germany.
Clinic for Plastic and Reconstructive Surgery, Aesthetic and Preventive Medicine at Heidelberg University Hospital-ETHIANUM, Heidelberg, Germany.
Cytotherapy. 2014 Jun;16(6):789-99. doi: 10.1016/j.jcyt.2014.01.007. Epub 2014 Mar 15.
Adipose tissue-derived stem cells (ADSCs) are thought to have great potential in regenerative medicine. A xenoprotein-free culture and handling system is desirable. To date, there is only little and contradictory information about the influence of the different types of human serum on ADSC proliferation and differentiation.
First, ADSCs were cultured in media containing regular human serum (HS plus) or fetal calf serum (FCS plus) with supplementation of growth factors for three passages. During passage 4, ADSC proliferative activity and adipogenic, osteogenic and chondrogenic differentiation ability was quantified. Second, ADSCs were cultured with three different human sera (regular human serum [HS], human serum from platelet-poor plasma [SPPP] or human serum from platelet-rich plasma [SPRP]) without supplementation of platelet-derived growth factor and assessed accordingly. The growth factor content of the different types of human sera was determined by means of multiplex protein assay and enzyme-linked immunosorbent assay.
The different sera did not affect ADSC doubling time significantly (P < 0.05). Specific glycerol-3-phosphat-dehydrogenase activity was significantly lower in cultures with SPRP (P < 0.01) compared with the other media compositions. Extracellular calcium deposition was significantly higher in cells differentiated in cultures with HS or SPPP compared with those with SPRP, HS plus or FCS (P < 0.01). Glycosaminoglycan content and collagen 2 were highest in cells cultured with SPRP (P < 0.001).
Culturing ADSCs in human serum appears to be a reasonable and efficient alternative compared with FCS. With respect to the outcome of a sighted clinical application, it appears to be feasible to handle the cells in a serum suitable for the intended later use.
脂肪组织来源的干细胞（ADSCs）在再生医学中具有巨大的潜力。理想的是使用无异种蛋白的培养和处理系统。迄今为止，关于不同类型的人血清对 ADSC 增殖和分化的影响，仅有少量且相互矛盾的信息。
首先，将 ADSC 培养在含有常规人血清（HS plus）或胎牛血清（FCS plus）的培养基中，并补充生长因子进行三传代。在传代 4 时，定量测定 ADSC 的增殖活性以及成脂、成骨和成软骨分化能力。其次，将 ADSC 分别用三种不同的人血清（常规人血清[HS]、血小板少血浆来源的人血清[SPPP]或血小板富血浆来源的人血清[SPRP]）培养，不补充血小板衍生生长因子，并进行相应评估。通过多重蛋白检测和酶联免疫吸附试验测定不同类型人血清中的生长因子含量。
不同的血清对 ADSC 倍增时间无显著影响（P<0.05）。与其他培养基成分相比，SPRP 中的甘油-3-磷酸脱氢酶活性明显较低（P<0.01）。在 HS 或 SPPP 培养的细胞中，细胞外钙沉积明显高于 SPRP 的细胞，在 HS 或 SPPP 培养的细胞中，细胞外钙沉积明显高于 SPRP、HS plus 或 FCS 的细胞（P<0.01）。在 SPRP 培养的细胞中，糖胺聚糖含量和胶原蛋白 2 最高（P<0.001）。
与 FCS 相比，在人血清中培养 ADSC 似乎是一种合理且高效的替代方法。就有目的的临床应用的结果而言，在适合预期后续用途的血清中处理细胞似乎是可行的。
