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Evaluation of platelet lysate as a substitute for FBS in explant and enzymatic isolation methods of human umbilical cord MSCs

机译:在人脐带间充质干细胞的外植体和酶分离方法中评估血小板裂解液替代FBS的价值

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摘要

Mesenchymal stem cells (MSCs) have immense potential for cell-based therapy of acute and chronic pathological conditions. MSC transplantation for cell-based therapy requires a substantial number of cells in the range of 0.5–2.5 × 106 cells/kg body weight of an individual. A prolific source of MSCs followed by in vitro propagation is therefore an absolute prerequisite for clinical applications. Umbilical cord tissue (UCT) is an abundantly available prolific source of MSC that are fetal in nature and have higher potential for ex-vivo expansion. However, the ex-vivo expansion of MSCs using a xenogeneic supplement such as fetal bovine serum (FBS) carries the risk of transmission of zoonotic infections and immunological reactions. We used platelet lysate (PL) as a xeno-free, allogeneic replacement for FBS and compared the biological and functional characteristics of MSC processed and expanded with PL and FBS by explant and enzymatic method. UCT-MSCs expanded using PL displayed typical immunophenotype, plasticity, immunomodulatory property and chromosomal stability. PL supplementation also showed 2-fold increase in MSC yield from explant culture with improved immunomodulatory activity as compared to enzymatically dissociated cultures. In conclusion, PL from expired platelets is a viable alternative to FBS for generating clinically relevant numbers of MSC from explant cultures over enzymatic method.
机译:间充质干细胞(MSCs)在基于细胞的急性和慢性病理状况治疗中具有巨大潜力。用于基于细胞的治疗的MSC移植需要大量的细胞,范围为0.5-2.5×10 6 个细胞/每公斤体重。因此,大量的MSC继之以体外繁殖是临床应用的绝对前提。脐带组织(UCT)是MSC的大量来源,其本质上是胎儿的,具有更高的离体扩增潜能。然而,使用异种补充剂如胎牛血清(FBS)进行的MSC的体外扩增具有传播人畜共患感染和免疫反应的风险。我们使用血小板裂解物(PL)作为FBS的无异种异体替代物,并通过外植体和酶促方法比较了用PL和FBS处理和扩增的MSC的生物学和功能特性。使用PL扩增的UCT-MSC具有典型的免疫表型,可塑性,免疫调节特性和染色体稳定性。 PL补充还显示,与酶解培养相比,外植体培养的MSC产量提高了2倍,具有增强的免疫调节活性。总之,来自过期血小板的PL是FBS的可行替代方案,可通过酶促方法从外植体培养物中产生临床相关数量的MSC。

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