TY - JOUR
T1 - In Vitro Culture Expansion Shifts the Immune Phenotype of Human Adipose-Derived Mesenchymal Stem Cells
AU - Jeske, Richard
AU - Yuan, Xuegang
AU - Fu, Qin
AU - Bunnell, Bruce A.
AU - Logan, Timothy M.
AU - Li, Yan
N1 - Funding Information:
This work was supported by National Science Foundation Award (CBET #1743426) and partially supported by the National Institutes of Health (NIH; R01NS102395). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Funding Information:
In fond memory, the authors acknowledge guidance provided by Professor Teng Ma. The authors also would like to thank Ms. Ruth Didier of FSU Department of Biomedical Sciences for her help in ?ow cytometry and Dr. Brian Washburn and Kristina Poduch of FSU Department of Biomedical Sciences for performing RT-PCR. In addition, the authors would like to thank Dr. Amber N. Brown for preparing the RNA libraries for transcriptome analysis, the Translational Science Laboratory at Florida State University for help in proteomics and mRNA-sequencing experiments, and Dr. Cynthia Vied for preliminary data analysis of mRNA-sequencing data.
Publisher Copyright:
© Copyright © 2021 Jeske, Yuan, Fu, Bunnell, Logan and Li.
PY - 2021/3/10
Y1 - 2021/3/10
N2 - Human mesenchymal stem or stromal cells (hMSCs) are known for their potential in regenerative medicine due to their differentiation abilities, secretion of trophic factors, and regulation of immune responses in damaged tissues. Due to the limited quantity of hMSCs typically isolated from bone marrow, other tissue sources, such as adipose tissue-derived mesenchymal stem cells (hASCs), are considered a promising alternative. However, differences have been observed for hASCs in the context of metabolic characteristics and response to in vitro culture stress compared to bone marrow derived hMSCs (BM-hMSCs). In particular, the relationship between metabolic homeostasis and stem cell functions, especially the immune phenotype and immunomodulation of hASCs, remains unknown. This study thoroughly assessed the changes in metabolism, redox cycles, and immune phenotype of hASCs during in vitro expansion. In contrast to BM-hMSCs, hASCs did not respond to culture stress significantly during expansion as limited cellular senescence was observed. Notably, hASCs exhibited the increased secretion of pro-inflammatory cytokines and the decreased secretion of anti-inflammatory cytokines after extended culture expansion. The NAD+/NADH redox cycle and other metabolic characteristics associated with aging were relatively stable, indicating that hASC functional decline may be regulated through an alternative mechanism rather than NAD+/Sirtuin aging pathways as observed in BM-hMSCs. Furthermore, transcriptome analysis by mRNA-sequencing revealed the upregulation of genes for pro-inflammatory cytokines/chemokines and the downregulation of genes for anti-inflammatory cytokines for hASCs at high passage. Proteomics analysis indicated key pathways (e.g., tRNA charging, EIF2 signaling, protein ubiquitination pathway) that may be associated with the immune phenotype shift of hASCs. Together, this study advances our understanding of the metabolism and senescence of hASCs and may offer vital insights for the biomanufacturing of hASCs for clinical use.
AB - Human mesenchymal stem or stromal cells (hMSCs) are known for their potential in regenerative medicine due to their differentiation abilities, secretion of trophic factors, and regulation of immune responses in damaged tissues. Due to the limited quantity of hMSCs typically isolated from bone marrow, other tissue sources, such as adipose tissue-derived mesenchymal stem cells (hASCs), are considered a promising alternative. However, differences have been observed for hASCs in the context of metabolic characteristics and response to in vitro culture stress compared to bone marrow derived hMSCs (BM-hMSCs). In particular, the relationship between metabolic homeostasis and stem cell functions, especially the immune phenotype and immunomodulation of hASCs, remains unknown. This study thoroughly assessed the changes in metabolism, redox cycles, and immune phenotype of hASCs during in vitro expansion. In contrast to BM-hMSCs, hASCs did not respond to culture stress significantly during expansion as limited cellular senescence was observed. Notably, hASCs exhibited the increased secretion of pro-inflammatory cytokines and the decreased secretion of anti-inflammatory cytokines after extended culture expansion. The NAD+/NADH redox cycle and other metabolic characteristics associated with aging were relatively stable, indicating that hASC functional decline may be regulated through an alternative mechanism rather than NAD+/Sirtuin aging pathways as observed in BM-hMSCs. Furthermore, transcriptome analysis by mRNA-sequencing revealed the upregulation of genes for pro-inflammatory cytokines/chemokines and the downregulation of genes for anti-inflammatory cytokines for hASCs at high passage. Proteomics analysis indicated key pathways (e.g., tRNA charging, EIF2 signaling, protein ubiquitination pathway) that may be associated with the immune phenotype shift of hASCs. Together, this study advances our understanding of the metabolism and senescence of hASCs and may offer vital insights for the biomanufacturing of hASCs for clinical use.
KW - NAD redox cycle
KW - adipose-derived mesenchymal stem cells
KW - immune phenotype
KW - proteomics
KW - replicative senescence
KW - transcriptomics
UR - http://www.scopus.com/inward/record.url?scp=85100436892&partnerID=8YFLogxK
U2 - 10.3389/fimmu.2021.621744
DO - 10.3389/fimmu.2021.621744
M3 - Article
C2 - 33777002
AN - SCOPUS:85100436892
SN - 1664-3224
VL - 12
JO - Frontiers in Immunology
JF - Frontiers in Immunology
M1 - 621744
ER -