Adipose tissue mesenchymal stem cell expansion in animal serum-free medium supplemented with autologous human platelet lysate
Adipose tissue mesenchymal stem cell expansion in animal serum-free medium supplemented with autologous human platelet lysate
复制标题
DOI：
10.1111/j.1537-2995.2009.02346.x
复制
发表时间：
2009-12-01
期刊：
影响因子：
2.9
通讯作者：
Krieger, Jose Eduardo
中科院分区：
文献类型：
--
作者：
Blande, Ivan Souza;Bassaneze, Vinicius;Krieger, Jose Eduardo
背景技术背景：间充质干细胞（MSC）已被认为是人类再生治疗应用，特别是在种间污染的情况下，需要安全的培养和扩增方案。人血小板裂解物（PL）已被提议作为MSC体外扩增过程中的动物血清替代物。在这项工作中，一个简单而有效的方法来获得自体PL，以取代动物血清在细胞培养applications.Study设计和方法：PL冷冻和离心程序获得的培养基补充人脂肪间充质干细胞（hASC）培养进行了测试。结果：与补充10%FBS相比，当用相同浓度的PL培养hASCs时，细胞群体倍增时间显著降低（PL 22.9 +/-1.5hr vs. FBS 106.7 +/-6.5hr，t检验，p < 0.05）。此外，用2.5%PL补充维持的hASC也显示出令人满意的结果。免疫表型分析显示，PL或FBS补充培养的hASC之间没有差异，两种培养物都保留了分化成脂肪细胞的潜力。这些结果表明，从同一供体获得的自体PL可用作动物血清替代品在hASC culture.CONCLUSIONS：合在一起，提供的证据表明，由单一供体提供的血小板足以获得PL的hASC繁殖临床规模的应用，减轻潜在的不良副作用与使用动物源性试剂。
BACKGROUND: Mesenchymal stem cells (MSCs) have been considered for human regenerative therapy applications, and safe culture and expansion protocols are needed especially in the context of interspecies contamination. Human platelet lysate (PL) has been proposed as animal serum substitute during in vitro MSC expansion. In this work, a simplified and efficient method to obtain autologous PL to replace animal serum in cell culture applications is described.STUDY DESIGN AND METHODS: PL obtained by freezing and centrifugation procedures was tested as medium supplement for human adipose mesenchymal stem cell (hASC) culture. Differential proliferation, immunophenotypic changes, and differentiation under PL or fetal bovine serum (FBS) were assessed.RESULTS: In contrast to 10% FBS supplementation, cell population doubling time was significantly lower when hASCs were cultured with the same concentration of PL ( PL 22.9 +/- 1.5 hr vs. FBS 106.7 +/- 6.5 hr, t test, p < 0.05). Furthermore, hASCs maintained with 2.5% PL supplementation also showed satisfactory results. Immunophenotypic analysis revealed no differences between hASCs cultivated with PL or FBS supplementation and both cultures retained the potential to differentiate into adipose cells. These results demonstrate that autologous PL obtained from the same donor can be used as animal serum substitute in hASC culture.CONCLUSIONS: Taken together, evidence is provided that platelets provided by a single donor are sufficient to obtain PL for hASC propagation for clinical-scale applications mitigating the potential untoward side effects associated with the use of animal-derived reagents.