Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue
Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue
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DOI：
10.1634/stemcells.2006-0627
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发表时间：
2007-01-01
期刊：
影响因子：
5.2
通讯作者：
Bieback, Karen
中科院分区：
文献类型：
--
作者：
Kocaoemer, Asli;Kern, Susanne;Bieback, Karen
目前，MSC在细胞治疗和组织工程中的临床潜力是关注的焦点。然而，大多数用于临床规模生产MSC的分离和扩增方案使用胎牛血清（FCS）作为补充，这造成了感染以及免疫反应的潜在风险。为了寻找合适的FCS替代物，我们以FCS作为标准对照培养基，研究了混合人AB血清（AB-HS）和凝血酶激活的富血小板血浆（tPRP）对脂肪组织间充质干细胞（AT-MSCs）的影响。在三种不同条件下培养10个供体的AT-MSC：（a）10%FCS，（B）10%ABHS，和（c）10%tPRP。沿着免疫表型，评估了集落形成单位、累积群体倍增率和向成脂和成骨谱系的分化能力。我们证明了AB-HS和tPRP对AT-MSCs的增殖作用显著高于FCS。在前六代中，AB-HS和tPRP MSC分别表现出66.6 +/- 15.7和68.1 +/- 6.7的倍数扩增，而FCS为24.4 +/- 0.7。分化能力在整个长期培养过程中得以保持。免疫表型是MSC的特征，并且在所有培养条件下具有可比性，除了AB-HS和tPRP的独特CD 45-/CD 14阳性侧群外，其倾向于随着培养时间的延长而减少。我们发现，合并的人AB血清和凝血酶活化的富血小板血浆是FCS用于AT-MSCs的替代品。这些人源在潜在感染威胁方面具有更好的特征，同时在整个长期培养中提供更高的增殖率并保持分化能力和间充质干细胞标志物表达。
MSCs are currently in focus regarding their clinical potential in cell therapy and tissue engineering. However, most isolation and expansion protocols for clinical-scale production of MSCs use fetal calf serum (FCS) as a supplement, which poses a potential risk for infections as well as immunological reactions. To find a suitable FCS substitute, we investigated the effects of pooled human AB serum (AB-HS) and thrombin-activated platelet-rich plasma (tPRP) on adipose tissue MSCs (AT-MSCs) with FCS as the standard control medium. AT-MSCs of 10 donors were cultured under three different conditions: (a) 10% FCS, (b) 10% ABHS, and (c) 10% tPRP. Colony-forming units, cumulative population doubling rates, and differentiation capacity toward the adipogenic and osteogenic lineages were assessed, along with immunophenotype. We demonstrated that AB-HS and tPRP provide a significantly higher proliferative effect on AT-MSCs than does FCS. In the first six passages, AB-HS and tPRP MSCs exhibited a fold expansion of 66.6 +/- 15.7 and 68.1 +/- 6.7, respectively, compared with 24.4 +/- 0.7 for FCS. Differentiation capacity was preserved throughout long-term culture. Immunophenotype was characteristic for MSCs and comparable for all culture conditions with the exception of a distinct CD45-/CD14-positive side population for AB-HS and tPRP that tended to diminish with prolonged culture. We showed that pooled human AB serum and thrombin-activated platelet-rich plasma are alternatives to FCS for AT-MSCs. These human sources are better characterized regarding potential infectious threats, while providing a higher proliferation rate and retaining differentiation capacity and mesenchymal stem cell marker expression throughout long-term culture.