Novel Methods to Mobilize, Isolate, and Expand Mesenchymal Stem Cells.

Cristiano P Vieira,Taralyn M Mccarrel,Maria B Grant

doi:10.3390/ijms22115728

Abstract

Numerous studies demonstrate the essential role of mesenchymal stem cells (MSCs) in the treatment of metabolic and inflammatory diseases, as these cells are known to modulate humoral and cellular immune responses. In this manuscript, we efficiently present two novel approaches to obtain MSCs from equine or human sources. In our first approach, we used electro-acupuncture as previously described by our group to mobilize MSCs into the peripheral blood of horses. For equine MSC collection, culture, and expansion, we used the Miltenyi Biotec CliniMACS Prodigy system of automated cell manufacturing. Using this system, we were able to generate appoximately 100 MSC colonies that exhibit surface marker expression of CD105 (92%), CD90 (85%), and CD73 (88%) within seven days of blood collection. Our second approach utilized the iPSC embryoid bodies from healthy or diabetic subjects where the iPSCs were cultured in standard media (endothelial + mesoderm basal media). After 21 days, the cells were FACS sorted and exhibited surface marker expression of CD105, CD90, and CD73. Both the equine cells and the human iPSC-derived MSCs were able to differentiate into adipogenic, osteogenic, and chondrogenic lineages. Both methods described simple and highly efficient methods to produce cells with surface markers phenotypically considered as MSCs and may, in the future, facilitate rapid production of MSCs with therapeutic potential.

Highlights

Our findings indicate that peripheral blood containing EA-mobilized Mesenchymal stem cells (MSCs), when processed using an automated system, gave superior results in which a larger number of MSC colonies expressed CD105, CD73, and CD90 by
We found that harvesting and differentiating embryoid bodies (EBs) from iPSCs with simplified culture media for 21 days resulted in the robust generation of MSCs from diabetic and healthy subjects with the same efficiency
Peripheral blood was obtained 3 h after the completion of the EA, a time previously optimized for the mobilization of MSCs

Summary

Introduction

Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Mesenchymal stem cells (MSCs) represent a versatile cell source for tissue regeneration [1] and remodeling due to their potent bioactivity, which includes modulation of inflammation, macrophage polarization toward pro-regenerative lineages, promotion of angiogenesis, and reduction in fibrosis. The paracrine signals of MSCs, their secretome, largely mediates their immunomodulatory and restorative potential [2]. There are close to 1000 clinical trials involving MSCs in the NIH clinical database (http://clinicaltrials.gov)

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