Division of Physiology, Innsbruck Medical University, Austria.
ALTEX. 2011;28(4):305-16. doi: 10.14573/altex.2011.4.305.
The search for alternatives to the use of fetal bovine serum (FBS) in cell and tissue culture media has become a major goal in terms of the 3R principles in order to reduce or to avoid harvesting of FBS from bovine fetuses, and, in terms of Good Manufacturing Practice (GMP), to ensure safe and animal product-free conditions for biomedical tissue engineering and (adult) stem cell therapy, respectively. In the present study, we investigated the feasibility of using platelet lysates (PL) as a substitute for FBS, based on the fact that most of the potent mitogenic factors present in serum are derived from activated thrombocytes. Platelet lysates were obtained from outdated human donor platelet concentrates. Methods were established to activate human donor platelets in order to achieve a maximum yield of platelet a-granule growth factors. Platelet lysates were successfully introduced to grow and maintain anchorage-dependent and -independent human and animal cell lines. For cell culture experiments, cells were either grown in culture media supplemented with 10% FBS, 5% PL, or under serum-free conditions. Growth experiments, viability assays, and platelet lysate-induced activation of ERK1/2 mitogen-activated protein kinase revealed platelet lysates as a valuable alternative to FBS in cell culture media.
寻找替代物来替代细胞和组织培养基中的胎牛血清(FBS)已成为 3R 原则的主要目标,以减少或避免从牛胎儿中提取 FBS,并且根据良好生产规范(GMP),确保用于生物医学组织工程和(成人)干细胞治疗的安全和无动物产品的条件。在本研究中,我们基于这样一个事实,即血清中存在的大多数有效的有丝分裂原因子均来自激活的血小板,研究了使用血小板裂解液(PL)作为 FBS 的替代品的可行性。血小板裂解液是从过期的人供体血小板浓缩物中获得的。建立了从人供体血小板中激活的方法,以实现血小板α-颗粒生长因子的最大产量。成功地将血小板裂解液引入以生长和维持锚定依赖性和非依赖性的人和动物细胞系。对于细胞培养实验,细胞要么在添加 10% FBS、5% PL 的培养基中生长,要么在无血清条件下生长。生长实验、活力测定和血小板裂解液诱导的 ERK1/2 丝裂原活化蛋白激酶的激活表明,血小板裂解液是细胞培养基中 FBS 的有价值替代品。