Improved Isolation of Mesenchymal Stem Cells Based on Interactions between N-Acetylglucosamine-Bearing Polymers and Cell-Surface Vimentin.

Hirohiko Ise,Yasuyuki Sakai,Kumiko Matsunaga,Marie Shinohara

doi:10.1155/2019/4341286

Hirohiko Ise, Yasuyuki Sakai + Show 2 more

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https://doi.org/10.1155/2019/4341286

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Abstract

Mesenchymal stem cells (MSCs) in bone marrow and adipose tissues are expected to be effective tools for regenerative medicine to treat various diseases. To obtain MSCs that possess both high differentiation and tissue regenerative potential, it is necessary to establish an isolation system that does not require long-term culture. It has previously been reported that the cytoskeletal protein vimentin, expressed on the surfaces of multiple cell types, possesses N-acetylglucosamine- (GlcNAc-) binding activity. Therefore, we tried to exploit this interaction to efficiently isolate MSCs from rat bone marrow cells using GlcNAc-bearing polymer-coated dishes. Cells isolated by this method were identified as MSCs because they were CD34-, CD45-, and CD11b/c-negative and CD90-, CD29-, CD44-, CD54-, CD73-, and CD105-positive. Osteoblast, adipocyte, and chondrocyte differentiation was observed in these cells. In total, yields of rat MSCs were threefold to fourfold higher using GlcNAc-bearing polymer-coated dishes than yields using conventional tissue-culture dishes. Interestingly, MSCs isolated with GlcNAc-bearing polymer-coated dishes strongly expressed CD106, whereas those isolated with conventional tissue-culture dishes had low CD106 expression. Moreover, senescence-associated β-galactosidase activity in MSCs from GlcNAc-bearing polymer-coated dishes was lower than that in MSCs from tissue-culture dishes. These results establish an improved isolation method for high-quality MSCs.

Highlights

Mesenchymal stem cells (MSCs) have the potential to differentiate into osteocytes, chondrocytes, adipocytes, hepatocytes, nerve cells, and vascular endothelial cells [1,2,3,4,5,6]
MSCs isolated with GlcNAc-bearing polymer-coated dishes strongly expressed CD106, whereas those isolated with conventional tissue-culture dishes had low CD106 expression
Seven types of AC-GlcNAc were produced with average molecular weights (Mw’s) of 2600–10300 Da and 8– 48 GlcNAc ligands (Figure 1(a) and Table 1)

Summary

Introduction

Mesenchymal stem cells (MSCs) have the potential to differentiate into osteocytes, chondrocytes, adipocytes, hepatocytes, nerve cells, and vascular endothelial cells [1,2,3,4,5,6] They are promising sources for regenerating cartilage and liver tissue [2,3,4,5,6,7]. MSCs produce multiple effective cytokines, growth factors, and exosomes that promote tissue regeneration [8, 9] These functions of MSCs tend to be unstable and difficult to maintain [10,11,12,13], likely as a result of long-term culture and high passage numbers. To obtain large numbers of MSCs without long-term culture, efficient isolation of MSCs with high proliferative activity is critical

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