Center of Orthopedics, Traumatology, and Spinal Cord Injury, Heidelberg University Hospital, Schlierbacher Landstraße 200a, 69118 Heidelberg, Germany.
Cells. 2020 Apr 9;9(4):918. doi: 10.3390/cells9040918.
Fetal calf serum (FCS) is frequently used as a growth factor and protein source in bone-marrow-derived mesenchymal stromal cell (BMSC) culture media, although it is a xenogenic product presenting multiple disadvantages including but not limited to ethical concerns. A promising alternative for FCS is human platelet lysate (hPL), which is produced out of human platelet concentrates and happens to be a stable and reliable protein source. In this study, we investigated the influence of hPL in an expansion medium (ESM) and an osteogenic differentiation medium (ODM) on the proliferation and osteogenic differentiation capacity of human BMSC. Therefore, we assessed population doublings during cell expansion, performed alizarin red staining to evaluate the calcium content in the extracellular matrix and determined the activity of alkaline phosphatase (ALP) as osteogenic differentiation correlates. The proliferation rate of BMSC cultured in ESM supplemented with hPL exceeded the proliferation rate of BMSC cultured in the presence of FCS. Furthermore, the calcium content and ALP activity was significantly higher in samples incubated in hPL-supplemented ODM, especially in the early phases of differentiation. Our results show that hPL can replace FCS as a protein supplier in cell culture media and does not negatively affect the osteogenic differentiation capacity of BMSC.
胎牛血清(FCS)常被用作骨髓间充质干细胞(BMSC)培养中的生长因子和蛋白来源,尽管它是一种异源产品,存在诸多缺点,包括但不限于伦理问题。人血小板裂解液(hPL)是 FCS 的一种有前途的替代品,它由人血小板浓缩物制成,恰好是一种稳定可靠的蛋白来源。在这项研究中,我们研究了 hPL 在扩展培养基(ESM)和成骨分化培养基(ODM)中的影响,以评估其对人 BMSC 的增殖和成骨分化能力的影响。因此,我们评估了细胞扩展过程中的倍增,通过茜素红染色评估细胞外基质中的钙含量,并测定碱性磷酸酶(ALP)的活性作为成骨分化的相关指标。在 hPL 补充的 ESM 中培养的 BMSC 的增殖速度超过了在 FCS 存在下培养的 BMSC 的增殖速度。此外,在 hPL 补充的 ODM 中孵育的样本的钙含量和 ALP 活性明显更高,尤其是在分化的早期阶段。我们的研究结果表明,hPL 可以替代 FCS 作为细胞培养基中的蛋白来源,并且不会对 BMSC 的成骨分化能力产生负面影响。