TY - JOUR
T1 - MicroRNA profiling reveals age-dependent differential expression of nuclear factor B and mitogen-activated protein kinase in adipose and bone marrow-derived human mesenchymal stem cells
AU - Pandey, Amitabh C.
AU - Semon, Julie A.
AU - Kaushal, Deepak
AU - O'Sullivan, Regina P.
AU - Glowacki, Julie
AU - Gimble, Jeffery M.
AU - Bunnell, Bruce A.
N1 - Funding Information:
We thank Alan Tucker for his excellent technical assistance in performing flow cytometry. We also thank Margie McCants and Christina McLennan for their technical assistance with cell culture and Dina Gaupp for her tremendous help with immunocytochemistry. We thank Drs. James Wade, Ann Reilley, and Michael Teague, together with their office staff and patients, for donating adipose tissues to this project. JMG is the Founder of LaCell, LLC, and serves as a consultant for Johnson & Johnson companies ATRM and Mentor. The research was supported by grant numbers RR00164 from the National Center for Research Resources, R21-NS059665 National Institutes of Neurological Disorders and Stroke, National Institutes of Health, Pennington Biomedical Research Center, and Tulane University.
PY - 2011
Y1 - 2011
N2 - Introduction. Mesenchymal stem cells (MSCs) play a central role in mediating endogenous repair of cell and tissue damage. Biologic aging is a universal process that results in changes at the cellular and molecular levels. In the present study, the role of microRNA (miRNA) in age-induced molecular changes in MSCs derived from adipose tissue (ASCs) and bone marrow (BMSCs) from young and old human donors were investigated by using an unbiased genome-wide approach. Methods. Human ASCs and BMSCs from young and old donors were cultured, and total RNA was isolated. The miRNA fraction was enriched and used to determine the expression profile of miRNA in young and old donor MSCs. Based on miRNA expression, differences in donor MSCs were further investigated by using differentiation assays, Western blot, immunocytochemistry, and bioinformatics. Results: Biologic aging demonstrated reduced osteogenic and adipogenic potential in ASCs isolated from older donors, whereas cell size, complexity, and cell-surface markers remained intact with aging. Analysis of miRNA profiles revealed that small subsets of active miRNAs changed secondary to aging. Evaluation of miRNA showed significantly decreased levels of gene expression of inhibitory kappa B kinase (IB), interleukin-1, inducible nitric oxide synthase (iNOS), mitogen-activated protein kinase/p38, ERK1/2, c-fos, and c-jun in MSCs from older donors by both bioinformatics and Western blot analysis. Nuclear factor kappa B (NF-B), myc, and interleukin-4 receptor mRNA levels were significantly elevated in aged cells from both the adipose and bone marrow depots. Immunocytochemistry showed nuclear localization in young donors, but a cytosolic predominance of phosphorylated NF-B in ASCs from older donors. Western blot demonstrated significantly elevated levels of NF-B subunits, p65 and p50, and AKT. Conclusions: These findings suggest that differential expression of miRNA is an integral component of biologic aging in MSCs.
AB - Introduction. Mesenchymal stem cells (MSCs) play a central role in mediating endogenous repair of cell and tissue damage. Biologic aging is a universal process that results in changes at the cellular and molecular levels. In the present study, the role of microRNA (miRNA) in age-induced molecular changes in MSCs derived from adipose tissue (ASCs) and bone marrow (BMSCs) from young and old human donors were investigated by using an unbiased genome-wide approach. Methods. Human ASCs and BMSCs from young and old donors were cultured, and total RNA was isolated. The miRNA fraction was enriched and used to determine the expression profile of miRNA in young and old donor MSCs. Based on miRNA expression, differences in donor MSCs were further investigated by using differentiation assays, Western blot, immunocytochemistry, and bioinformatics. Results: Biologic aging demonstrated reduced osteogenic and adipogenic potential in ASCs isolated from older donors, whereas cell size, complexity, and cell-surface markers remained intact with aging. Analysis of miRNA profiles revealed that small subsets of active miRNAs changed secondary to aging. Evaluation of miRNA showed significantly decreased levels of gene expression of inhibitory kappa B kinase (IB), interleukin-1, inducible nitric oxide synthase (iNOS), mitogen-activated protein kinase/p38, ERK1/2, c-fos, and c-jun in MSCs from older donors by both bioinformatics and Western blot analysis. Nuclear factor kappa B (NF-B), myc, and interleukin-4 receptor mRNA levels were significantly elevated in aged cells from both the adipose and bone marrow depots. Immunocytochemistry showed nuclear localization in young donors, but a cytosolic predominance of phosphorylated NF-B in ASCs from older donors. Western blot demonstrated significantly elevated levels of NF-B subunits, p65 and p50, and AKT. Conclusions: These findings suggest that differential expression of miRNA is an integral component of biologic aging in MSCs.
UR - http://www.scopus.com/inward/record.url?scp=84857802021&partnerID=8YFLogxK
U2 - 10.1186/scrt90
DO - 10.1186/scrt90
M3 - Article
C2 - 22169120
AN - SCOPUS:84857802021
SN - 1757-6512
VL - 2
JO - Stem Cell Research and Therapy
JF - Stem Cell Research and Therapy
IS - 6
M1 - 49
ER -