Gottipamula Sanjay, Ashwin K M, Muttigi Manjunatha S, Kannan Suresh, Kolkundkar Udaykumar, Seetharam Raviraja N
Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
Cell Tissue Res. 2014 Apr;356(1):123-35. doi: 10.1007/s00441-013-1783-7. Epub 2014 Jan 22.
Bone marrow-derived mesenchymal stromal cells (BM-MSCs) heralded a new beginning for regenerative medicine and generated tremendous interest as the most promising source for therapeutic application. Most cell therapies require stringent regulatory compliance and prefer the use of serum-free media (SFM) or xeno-free media (XFM) for the MSC production process, starting from the isolation onwards. Here, we report on serum-free isolation and expansion of MSCs and compare them with cells grown in conventional fetal bovine serum (FBS)-containing media as a control. The isolation, proliferation and morphology analysis demonstrated significant differences between MSCs cultured in various SFM/XFM in addition to their difference with FBS controls. BD Mosaic™ Mesenchymal Stem Cell Serum-Free media (BD-SFM) and Mesencult-XF (MSX) supported the isolation, sequential passaging, tri-lineage differentiation potential and acceptable surface marker expression profile of BM-MSCs. Further, MSCs cultured in SFM showed higher immune suppression and hypo-immunogenicity properties, making them an ideal candidate for allogeneic cell therapy. Although cells cultured in control media have a significantly higher proliferation rate, BM-MSCs cultured in BD-SFM or MSX media are the preferred choice to meet regulatory requirements as they do not contain bovine serum. While BM-MSCs cultured in BD-SFM and MSX media adhered to all MSC characteristics, in the case of few parameters, the performance of cells cultured in BD-SFM was superior to that of MSX media. Pre-clinical safety and efficiency studies are required before qualifying SFM or XFM media-derived MSCs for therapeutic applications.
骨髓间充质基质细胞（BM-MSCs）为再生医学开启了新的篇章，并作为最具前景的治疗应用来源引发了极大的关注。大多数细胞疗法需要严格遵守监管规定，并且从分离开始，在MSC生产过程中更倾向于使用无血清培养基（SFM）或无异源培养基（XFM）。在此，我们报告了MSCs的无血清分离和扩增，并将其与在传统含胎牛血清（FBS）培养基中培养的细胞作为对照进行比较。分离、增殖和形态学分析表明，在各种SFM/XFM中培养的MSCs之间存在显著差异，此外它们与FBS对照也存在差异。BD Mosaic™间充质干细胞无血清培养基（BD-SFM）和Mesencult-XF（MSX）支持BM-MSCs的分离、连续传代、三系分化潜能以及可接受的表面标志物表达谱。此外，在SFM中培养的MSCs表现出更高的免疫抑制和低免疫原性特性，使其成为同种异体细胞治疗的理想候选者。尽管在对照培养基中培养的细胞增殖率显著更高，但在BD-SFM或MSX培养基中培养的BM-MSCs是满足监管要求的首选，因为它们不含牛血清。虽然在BD-SFM和MSX培养基中培养的BM-MSCs符合所有MSC特征，但在少数参数方面，在BD-SFM中培养的细胞性能优于MSX培养基。在将SFM或XFM培养基来源的MSCs用于治疗应用之前，需要进行临床前安全性和有效性研究。
