Department of Urology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
Biomed Eng Online. 2013 Feb 20;12:15. doi: 10.1186/1475-925X-12-15.
The conventional expansion of human mesenchymal stromal cells (hMSC) for tissue engineering or (pre-) clinical investigation includes the use of 10% fetal bovine serum (FBS). However, there exists immense lot-to-lot variability in FBS samples and time consuming as well as cost intensive lot pre-testing is essential to guarantee optimal hMSC proliferation and stem cells characteristics maintenance. Furthermore, lot-to-lot variability impedes the long-term consistency of research and comparability between research groups. Therefore, we investigated the use of defined, invariable, non-synthetic FBS in low serum culture conditions for isolation and expansion of hMSC.
hMSC were isolated from bone marrow in Panserin 401 supplemented with growth factors and 2% MSC-tested or non-tested, defined, invariable, non-synthetic FBS and further cultivated in vitro. The surface marker expression, differentiation capacity as well as cell proliferation and cytotoxicity was analyzed and compared between serum samples.
Cells isolated and cultivated with low concentrations of MSC-tested or non-tested FBS demonstrated no differences in surface marker expression or differentiation capacity. Proliferation of hMSC was equal in medium supplemented with either serum with no indication of cell death.
The low serum concentration in Panserin 401 supplemented with growth factors enables the use of defined, invariable, non-synthetic FBS for the isolation and expansion of hMSC. The required hMSC characteristics like surface marker expression and differentiation capacity are maintained. Importantly, no differences in the cell proliferation could be detected. Therefore, using these low-serum culture conditions, the need for lot-to-lot pre-testing of FBS usually needed for optimal hMSC expansion is abolished leading to long-term consistency and comparability of results.
用于组织工程或（临床前）研究的人源间充质基质细胞（hMSC）的常规扩增包括使用 10%胎牛血清（FBS）。然而，FBS 样本存在巨大的批次间变异性，需要进行耗时且成本高昂的批前测试，以保证 hMSC 的最佳增殖和干细胞特性维持。此外，批次间变异性阻碍了研究的长期一致性和研究小组之间的可比性。因此，我们研究了在低血清培养条件下使用定义的、不变的、非合成的 FBS 分离和扩增 hMSC 的用途。
hMSC 从骨髓中分离出来，在 Panserin 401 中添加生长因子，并使用 2%经过 MSC 测试或未经测试的定义的、不变的、非合成的 FBS 进行分离和扩增，并在体外进一步培养。分析和比较了血清样本之间的表面标志物表达、分化能力以及细胞增殖和细胞毒性。
用低浓度的经过 MSC 测试或未经测试的 FBS 分离和培养的细胞在表面标志物表达或分化能力方面没有差异。在补充有血清的培养基中 hMSC 的增殖是相等的，没有细胞死亡的迹象。
在 Panserin 401 中添加生长因子的低血清浓度允许使用定义的、不变的、非合成的 FBS 分离和扩增 hMSC。维持所需的 hMSC 特性，如表面标志物表达和分化能力。重要的是，细胞增殖没有差异。因此，使用这些低血清培养条件，消除了通常需要进行批前测试以实现最佳 hMSC 扩增的 FBS 的需求，从而实现结果的长期一致性和可比性。
