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Stepwise preconditioning enhances mesenchymal stem cell-based cartilage regeneration through epigenetic modification.

Abstract：

OBJECTIVE:This study is to investigate the functions and underlying mechanisms of mesenchymal stem cells (MSCs) underwent stepwise preconditioning in chondrogenic medium before expansion, then further explore their therapeutic effects in a surgically induced osteoarthritis (OA) model. METHODS:MSCs isolated from the adult rats expressing Green Fluorescence Protein (GFP) were incubated in basal medium or primed in chondrogenic medium before expansion. The multipotency including cell proliferation, differentiation, and survivability was compared between chondrogenic manipulated MSCs (M-MSCs) and untreated MSCs. Methylation modification of Nanog and Oct4 were detected by bisulfite genomic sequencing. Loss-of-function phenotype in M-MSCs induced by shNanog was also observed. Then the therapeutic effect of the cells was evaluated in a surgically induced OA rat model by single intraarticular injection. The injected GFP-labeled cells in the joints were monitored in vivo. These rats were sacrificed and subjected to histological examinations and microstructural analysis after 4 weeks. RESULTS:We found that cell clonogenicity, proliferation, survivability, and chondrogenic property were enhanced after stepwise preconditioning. We then further found that the expression level of Nanog and Oct4 was temporarily increased in the M-MSCs. Results of epigenetic analysis revealed that demethylation happened in Nanog and Oct4 after the stepwise preconditioning. Results of in vivo imaging showed more GFP-labeled cells in the M-MSCs-injected group. And results of histology and micro-CT analysis also indicated a superior therapeutic effect of M-MSCs on the surgically induced-OA. CONCLUSION:These findings indicated a feasible method to obtain a cell population with high survivability and chondrogenic commitment for the treatment of OA.

journal_name

Osteoarthritis Cartilage

journal_title

Osteoarthritis and cartilage

authors

Lin S,Lee WYW,Xu L,Wang Y,Chen Y,Ho KKW,Qin L,Jiang X,Cui L,Li G

doi

10.1016/j.joca.2017.05.008

subject

Has Abstract

pub_date

2017-09-01 00:00:00

pages

1541-1550

issue

9

eissn

1063-4584

issn

1522-9653

pii

S1063-4584(17)30995-0

journal_volume

25

pub_type

杂志文章

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