Serum-free process development: improving the yield and consistency of human mesenchymal stromal cell production

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• 作者：Thomas R.J. Heathman¹ ; Alexandra Stolzing¹ ; Claire Fabian² ; ³ ; Qasim A. Rafiq¹ ; ⁴ ; Karen Coopman¹ ; Alvin W. Nienow¹ ; ⁵ ; Bo Kara⁶ ; Christopher J. Hewitt¹ ; ⁴ ; ^{c.j.hewitt@aston.ac.uk" class="auth_mail" title="E-mail the corresponding author}
• 关键词：cell-based therapy ; comparability ; consistency ; human mesenchymal stromal cell ; manufacturing ; regenerative medicine ; serum-free ; yield
• 刊名：Cytotherapy
• 出版年：2015
• 出版时间：November 2015
• 年：2015
• 卷：17
• 期：11
• 页码：1524-1535
• 全文大小：1337 K

文摘

The cost-effective production of human mesenchymal stromal cells (hMSCs) for off-the-shelf and patient specific therapies will require an increasing focus on improving product yield and driving manufacturing consistency.

Methods

Bone marrow–derived hMSCs (BM-hMSCs) from two donors were expanded for 36 days in monolayer with medium supplemented with either fetal bovine serum (FBS) or PRIME-XV serum-free medium (SFM). Cells were assessed throughout culture for proliferation, mean cell diameter, colony-forming potential, osteogenic potential, gene expression and metabolites.

Results

Expansion of BM-hMSCs in PRIME-XV SFM resulted in a significantly higher growth rate (P < 0.001) and increased consistency between donors compared with FBS-based culture. FBS-based culture showed an inter-batch production range of 0.9 and 5 days per dose compared with 0.5 and 0.6 days in SFM for each BM-hMSC donor line. The consistency between donors was also improved by the use of PRIME-XV SFM, with a production range of 0.9 days compared with 19.4 days in FBS-based culture. Mean cell diameter has also been demonstrated as a process metric for BM-hMSC growth rate and senescence through a correlation (R² = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, in vitro colony-forming potential and osteogenic potential despite the higher number of population doublings.

Conclusions

We have increased the yield and consistency of BM-hMSC expansion between donors, demonstrating a level of control over the product, which has the potential to increase the cost-effectiveness and reduce the risk in these manufacturing processes.