Wu Xiaoyun, Kang Huiyan, Liu Xuemin, Gao Jin, Zhao Kuijun, Ma Zhijie
Department of pharmacy, Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Department of Technology, Beijing JingMeng Stem Cell Technology. Co. Ltd., Beijing, China.
Cell Prolif. 2016 Oct;49(5):579-88. doi: 10.1111/cpr.12279. Epub 2016 Aug 4.
Umbilical cord mesenchymal stromal cells (UCMSCs) can be considered to become a new gold standard for MSC-based therapies. A serum and xeno-free, chemically defined and no-plate-coating-based culture system will greatly facilitate development of robust, clinically acceptable bioprocesses for reproducibly generating quality-assured UCMSCs.
In this study, we report for the first time, such a serum-free, xeno-free, completely chemically defined and no-plate-coating-based culture system for the isolation and expansion of UCMSCs, whose biological characteristics were evaluated and compared with serum-containing medium (SCM) methods.
This culture system not only supported UCMSC primary cultures but also allowed for their expansion at low seeding density. Compared to SCM, UCMSCs in SFM exhibited (i) higher proliferative and colony-forming capacities; (ii) distinctly different morphologies; (iii) similar phenotype; (iv) similar pluripotency-associated marker expression; (v) superior osteogenic, but reduced adipogenic differentiation capacitities. In addition, UCMSCs cultured in SFM retained similar immunomodulatory properties to those in SCM.
Our findings demonstrate the feasibility of isolating and expanding UCMSCs in a completely serum-free, xeno-free, chemically defined and no-plate-coating-based culture system and represent an important step forward for development of robust, clinically acceptable bioprocesses for UCMSCs. Further, this provides a superior study platform for UCMSCs biology in a controlled environment.
脐带间充质基质细胞(UCMSCs)可被视为基于间充质干细胞疗法的新金标准。一种无血清、无异种成分、化学成分明确且无需包被培养板的培养系统将极大地促进稳健、临床可接受的生物工艺的发展,以可重复地生产质量有保证的UCMSCs。
在本研究中,我们首次报道了这样一种无血清、无异种成分、完全化学成分明确且无需包被培养板的培养系统,用于UCMSCs的分离和扩增,并对其生物学特性进行了评估,并与含血清培养基(SCM)方法进行了比较。
该培养系统不仅支持UCMSC原代培养,还能在低接种密度下实现其扩增。与SCM相比,SFM中的UCMSCs表现出:(i)更高的增殖和集落形成能力;(ii)明显不同的形态;(iii)相似的表型;(iv)相似的多能性相关标志物表达;(v)更强的成骨分化能力,但脂肪生成分化能力降低。此外,在SFM中培养的UCMSCs与在SCM中培养的UCMSCs具有相似的免疫调节特性。
我们的研究结果证明了在完全无血清、无异种成分、化学成分明确且无需包被培养板的培养系统中分离和扩增UCMSCs的可行性,这是开发稳健、临床可接受的UCMSCs生物工艺的重要一步。此外,这为在可控环境中研究UCMSCs生物学提供了一个优越的平台。