Stempeutics Research Pvt. Ltd, Shirdi Sai Baba Cancer Hospital, Manipal, India.
Biotechnol Lett. 2012 Jul;34(7):1367-74. doi: 10.1007/s10529-012-0893-8. Epub 2012 Apr 4.
Human platelet lysate (HPL) was evaluated as an alternative to fetal bovine serum (FBS) in large-scale culturing of bone marrow-derived mesenchymal stromal cells (BM-MSCs) for therapeutic applications. Dulbecco's modified Eagle medium (DMEM)of low glucose (LG) and Knock Out (KO) were used with human platelet lysate (HPL) as LG-HPL and KO-HPL, and with FBS as LG-FBS and KO-FBS to culture the BM-MSCs. HPL at 10 % (v/v) supported BM-MSCs growth and subsequent isolation efficiency generated >90 × 10(6) MSCs in LG-HPL. Population doublings (PDs) and population doubling times of LG-HPL and KO-HPL (PDT) were not significantly different but LG-HPL showed a significant clonogenic potential and HPL cultures had an average PDT of 36.5 ± 6.5 h and an average PDs of 5 ± 0.7/passage. BM-MSCs cultured with LG-HPL had significantly higher immunosuppression compared to LG-FBS, but KO-HPL and KO-FBS-grown cultures were not significantly different. HPL is therefore alternative to FBS for large-scale production of BM-MSCs for therapeutic applications.
人血小板裂解液(HPL)被评估为替代胎牛血清(FBS),用于治疗应用中大规模培养骨髓间充质基质细胞(BM-MSCs)。低葡萄糖(LG)和无蛋白(KO)的 Dulbecco改良 Eagle 培养基(DMEM)与 HPL 一起使用,作为 LG-HPL 和 KO-HPL,与 FBS 一起作为 LG-FBS 和 KO-FBS 来培养 BM-MSCs。10%(v/v)的 HPL 支持 BM-MSCs 的生长,随后的分离效率在 LG-HPL 中产生了超过 90×10^6^个 MSC。LG-HPL 和 KO-HPL 的群体倍增(PD)和群体倍增时间(PDT)没有显著差异,但 LG-HPL 显示出显著的集落形成潜力,HPL 培养物的平均 PDT 为 36.5±6.5 h,平均 PDs 为 5±0.7/传代。与 LG-FBS 相比,用 LG-HPL 培养的 BM-MSCs 具有显著更高的免疫抑制作用,但 KO-HPL 和 KO-FBS 培养物之间没有显著差异。因此,HPL 是替代 FBS 用于治疗应用中 BM-MSCs 大规模生产的选择。