Institute for Neuropathology, Aachen University Hospital, RWTH Aachen, Germany.
Cytotherapy. 2010 Sep;12(5):587-92. doi: 10.3109/14653240903505814.
Human mesenchymal stromal cells (hMSC) are of enormous interest for various clinical applications. For the expansion of isolated hMSC to relevant numbers for clinical applications, 10% fetal bovine serum (FBS)-supplemented medium is commonly used. The main critical disadvantage of FBS is the possibility of transmission of infectious agents as well as the possibility of immune rejection of the transplanted cells in response to the bovine serum. Therefore, we tested a commercially available medium, Panserin 401, that was specifically developed for serum-free cell cultivation.
hMSC were isolated from bone marrow (BM) and expanded in either Dulbecco's modified Eagle medium (DMEM) or Panserin 401 alone, or combined with FBS (2% or 10%), with or without supplementary growth factors. Cell proliferation and cytotoxicity were monitored twice a week for 3 weeks.
No proliferation was observed in any of the serum-free media. However, DMEM/10% FBS (the conventional culture medium for hMSC) and DMEM/2% FBS with growth factors revealed moderate proliferation. Interestingly, the best proliferation was obtained using Panserin 401 supplemented with 2% FBS and growth factors (as well as with 10% FBS). Analysis of cell growth in Panserin 401 supplemented with 2% FBS only or with growth factors only revealed no proliferation, demonstrating the necessity of the combination of 2% FBS and growth factors. Efficient isolation and expansion of hMSC from cancellous bone could also be performed using Panserin 401 with 2% FBS and growth factors. Furthermore, these isolated cultures maintained multipotency, as demonstrated by adipogenic and osteogenic differentiation.
人骨髓间充质基质细胞（hMSC）对于各种临床应用具有巨大的兴趣。为了将分离的 hMSC 扩增到用于临床应用的相关数量，通常使用添加 10%胎牛血清（FBS）的培养基。FBS 的主要关键缺点是存在传染因子的传播可能性，以及移植细胞对牛血清的免疫排斥反应的可能性。因此，我们测试了一种商业上可获得的培养基，即专门为无血清细胞培养开发的 Panserin 401。
hMSC 从骨髓（BM）中分离出来，并在单独的 Dulbecco 改良 Eagle 培养基（DMEM）或 Panserin 401 中，或在添加 2%或 10% FBS 以及补充生长因子的情况下进行扩增。每周两次监测细胞增殖和细胞毒性，持续 3 周。
在任何无血清培养基中都没有观察到增殖。然而，DMEM/10% FBS（hMSC 的常规培养基）和 DMEM/2% FBS 与生长因子显示出适度的增殖。有趣的是，使用补充有 2% FBS 和生长因子的 Panserin 401 可获得最佳的增殖（以及使用 10% FBS）。仅用 2% FBS 或仅用生长因子补充 Panserin 401 时细胞生长的分析表明没有增殖，这表明 2% FBS 和生长因子的组合是必要的。使用补充有 2% FBS 和生长因子的 Panserin 401 也可以从松质骨中有效地分离和扩增 hMSC。此外，这些分离的培养物保持了多能性，如脂肪生成和成骨分化所证明的那样。
