Pharmaceutical Production Research Facility, Schulich School of Engineering, University of Calgary, 2500 University Drive NW, Calgary, Alberta, Canada.
J Tissue Eng Regen Med. 2012 May;6(5):391-403. doi: 10.1002/term.441. Epub 2011 Jul 11.
Human mesenchymal stem cells (hMSCs) are typically obtained for research or therapeutic applications by isolating and subculturing adherent cells from bone marrow on tissue-culture substrates using growth media. The purity and properties of the resulting populations can be affected greatly by the conditions under which they are cultured. Fetal bovine serum (FBS), although ill-defined, has been widely used as a critical requirement for conventional hMSC culture. However, a defined serum-free medium would greatly facilitate the development of robust, clinically acceptable bioprocesses for reproducibly generating quality-assured cells. The present study provides evidence demonstrating that a defined serum-free medium (PPRF-msc6) shows several beneficial features over a conventional FBS-containing medium for the production of hMSCs. When compared to control FBS-based cultures, PPRF-msc6 medium supported the derivation of hMSCs from primary cultures of bone marrow cells in a more rapid and consistent manner. Furthermore, hMSCs cultured in PPRF-msc6 exhibited: (a) a greater colony-forming capacity in primary as well as passaged cultures; (b) negligible lag phase and explicit exponential growth; (c) lower population doubling times (21-26 h vs 35-38 h between passage levels 1 and 10); (d) a greater number of population doublings (62 ± 4 vs 43 ± 2; over a 2 month period); and (e) a higher degree of homogeneity in size. Our data demonstrate that PPRF-msc6 is an important development which opens the door for the rapid, efficient and reproducible production of hMSCs in clinical settings.
人骨髓间充质干细胞（hMSCs）通常通过在组织培养基质上从骨髓中分离和传代贴壁细胞，并使用生长培养基来获得，用于研究或治疗应用。培养条件极大地影响所得细胞群体的纯度和特性。胎牛血清（FBS）虽然定义不明确，但已被广泛用作传统 hMSC 培养的关键要求。然而，无血清的定义培养基将极大地促进稳健、可临床接受的生物工艺的发展，以可重复地生成质量保证的细胞。本研究提供的证据表明，与传统含 FBS 的培养基相比，无血清定义培养基（PPRF-msc6）在 hMSC 的生产中具有几个有益的特性。与基于对照 FBS 的培养物相比，PPRF-msc6 培养基以更快速且更一致的方式支持从骨髓细胞的原代培养物中衍生出 hMSCs。此外，在 PPRF-msc6 中培养的 hMSCs表现出：（a）在原代和传代培养物中具有更大的集落形成能力；（b）可忽略的迟滞期和明确的指数生长；（c）更低的群体倍增时间（第 1 代和第 10 代之间的 21-26 小时与 35-38 小时）；（d）更高的群体倍增次数（2 个月期间的 62 ± 4 与 43 ± 2 次）；和（e）更大的大小均匀度。我们的数据表明，PPRF-msc6 是一个重要的发展，为在临床环境中快速、高效且可重复地生产 hMSCs 开辟了道路。
