Age-associated changes in the transcriptomes of non-cultured adipose-derived stem cells from young and old mice assessed via single-cell transcriptome analysis.

Yuta Doshida,Masayuki Yoshida,Haruka Sano,Toshiro Aigaki,Shinichi Hashimoto,Sadahiro Iwabuchi,Akihito Ishigami

doi:10.1371/journal.pone.0242171

Abstract

Adipose-derived stem cells (ASCs) exhibit self-renewal and pluripotency. The differentiation potency of ASCs has been reported to deteriorate with aging; however, relevant studies used ASCs that were isolated and subcultured several times. It is still unclear whether subcultured ASCs accurately reflect the in vivo state. To address this question, we used freshly isolated stromal vascular fractions (SVFs) and performed comprehensive single-cell transcriptome analysis. In this study, we identified three cell populations as putative ASC candidates in SVFs and three novel ASC-related genes: Adamts7, Snai2, and Tgfbr1, that are reported to be negative regulators of cell differentiation. Moreover, we identified age-associated high gene expression levels of Adamts7, Egfr, and Igfbp4 in the earliest differentiation stage of ASCs. These results suggest that aging may make it impossible to maintain the stringency of the regulation of the expression of some genes related to ASC differentiation.

Highlights

Adipose-derived stem cells (ASCs) are highly convenient multipotent mesenchymal stem cells for clinical application in human regenerative medicine because they are isolated from adipose tissues [1,2,3,4]
The slides were covered with a dialysis membrane and incubated with a cell lysis solution containing detergents
stromal vascular fraction (SVF) from mouse epididymal white adipose tissues were isolated by collagenase digestion and centrifugation to remove low-density fatty mature adipocytes, as described in the Methods and illustrated in S1 Fig. The isolated SVFs were subjected to our previously developed single-cell transcriptome

Summary

Introduction

Adipose-derived stem cells (ASCs) are highly convenient multipotent mesenchymal stem cells for clinical application in human regenerative medicine because they are isolated from adipose tissues [1,2,3,4]. Isolated ASCs can be stably subcultured in large quantities at the same time and be induced to differentiate into adipogenic, osteogenic, chondrogenic, and angiogenic cells [3]. Many patients who receive regenerative medicine are elderly people, so the isolation of ASCs from adipose tissue is likely to be more common in elderly people. It is very important to ensure that ASCs from elderly people are comparable to those from young people. Transcriptomes in non-cultured ASCs with age Competing interests: The authors declare no conflicts of interest

Methods

Results

Discussion

Conclusion