A therapy-grade protocol for differentiation of pluripotent stem cells into mesenchymal stem cells using platelet lysate as supplement

使用血小板裂解物作为补充剂将多能干细胞分化为间充质干细胞的治疗级方案

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作者:Carlos Luzzani, Gabriel Neiman, Ximena Garate, María Questa, Claudia Solari, Darío Fernandez Espinosa, Marcela García, Ana Lía Errecalde, Alejandra Guberman, María Elida Scassa, Gustavo Emilio Sevlever, Leonardo Romorini, Santiago Gabriel Miriuka

Conclusions

This protocol can be used to generate a large amount of PD-MSCs with low cost and is compatible with clinical therapies.

Methods

In this work we present a therapy-grade, inexpensive and simple protocol to derive MSCs from pluripotent stem cells (PSCs) based on the use of platelet lysate (PL) as medium supplement.

Results

We showed that the PD-MSCPL expressed multiple MSC markers, including CD90, CD73, CD105, CD166, and CD271, among others. These cells also show multilineage differentiation ability and immunomodulatory effects on pre-stimulated lymphocytes. Thorough characterization of these cells showed that a PD-MSCPL resembles an umbilical cord (UC) MSC and differs from a PSC in surface marker and extracellular matrix proteins and integrin expression. Moreover, the OCT-4 promoter is re-methylated with mesenchymal differentiation comparable with the methylation levels of UC-MSCs and fibroblasts. Lastly, the use of PL-supplemented medium generates significantly more MSCs than the use of fetal bovine serum. Conclusions: This protocol can be used to generate a large amount of PD-MSCs with low cost and is compatible with clinical therapies.

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