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Trichostatin A promotes cardiomyocyte differentiation of rat mesenchymel stem cells after 5-azacytidine induction or during co-culture with neonatal cardiomyocytes via a mechanism independent of histone deacetylase inhibition
Authors: Ge Yang, Jie Tian, Chuan Feng, Li-li Zhao, Zhenguo Liu, Jing Zhu
Source: Cell Transplantation
Publisher: Cognizant Communication Corporation
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Abstract:
This study was to investigate the effect of trichostatin A (TSA), a histone deacetylase (HDAC) inhibitor, on cardiac differentiation of bone marrow mesenchymal stem cells (MSCs) in vitro. Rat MSCs were isolated and divided into 6 groups: 1) control; 2) 5-azacytidine treatment (5-aza, 10 μM); 3) treatment with TSA (100, 300, and 500 nM); 4) treatment with 5-aza followed by incubation with TSA; 5) co-culture with neonatal cardiomyocytes (CMs) ; and 6) treatment with TSA then co-culture with CMs. HDAC activity was significantly inhibited in TSA-treated cells with the maximal inhibition after 24 hrs of exposure to TSA at 300 nM. No changes in HDAC activity were observed in control, 5-aza-treated or co-culture groups. Following 7 days of differentiation, the expression of early cardiac transcription factors GATA-4, NKx2.5, and MEF2c, cardiac troponin T (cTnT) was increased by 6-8 times in the cells in 5-aza-treated, co-culture, or TSA-treated groups over control as determined using real-time PCR, immunofluorescence staining, and Western blotting. However, the percent cTnT-positive cells dramatically different with 0.7% for control, 10% for 5-aza-treated, 25% for co-culture, and 4% for TSA-treated group (500 nM). TSA treatment of the cells pre-treated with 5-aza or co-cultured with CMs dramatically increased the expression of GATA-4, NKx2.5, and MEF2c by 35-50 times over control. The cTnT protein expression was also significantly increased by over 3-fold by TSA treatment (500 nM) in both 5-aza-treated and co-culture group over control. The percent cTnT-positive cells in both 5-aza-pre-treated and co-culture groups were significantly increased by TSA treatment after one week of differentiation by up to 92.6% (from 10.3% to 19.8%) and 23.9% (from 24.5% to 30.2%), respectively. These data suggested that TSA enhanced the cardiac differentiation of MSCs after 5-aza induction or during co-culture with CMs through a mechanism beyond the inhibition of HDAC activity.Document Type:
DOI: http://dx.doi.org/10.3727/096368911X593145
Appeared or available online: September 22, 2011
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