Feasibility of cardiac differentiation of bone marrow mesenchymal stem cells induced by co-culture indirectly with cardiomyocytes in several dimensions

doi:10.3724/SP.J.1123.2016.01006

Abstract

Abstract:

The objective of this research is to investigate the feasibility of cardiac differentiation of bone marrow mesenchymal stem cells (MSCs) by co-culture with cardiomyocytes (CMs) in vitro. The third generation of MSCs from bone marrow and CMs were co-cultured indirectly in a transwell. One week later, the expressions of muscle-specific markers (cardiac troponin I and α-actin) by immunofluorescence staining and the gene expressions of transcription factors (Nkx-2.5 and GATA-4) were measured by real-time polymerase chain reaction (RT-PCR). Then, orthogonal partial least squares discriminant analysis (OPLS-DA) models were employed to confirm the difference among MSCs, induced MSCs and CMs by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis. The distinctive changes were identified by multivariate analysis and variate analysis, and the changed metabolites were identified by MS and MS/MS. One week after co-cultured with CMs indirectly, the specific myocardium morphology of MSCs was observed under microscope. The positive immunofluorescence stainings against cTnI and α-actin were detected, and the positive expressions of the transcription factors Nkx-2.5 and GATA-4 were measured by RT-PCR. In OPLS-DA mode, obvious trend of cardiac differentiation of MSCs can be seen, and seven metabolites were tested both in induced MSCs and CMs, but five metabolites were tested in induced MSCs or CMs. Cardiac differentiation of MSCs can be induced by co-cultured with CMs indirectly in vitro. However, metabolism difference still existed between induced MSCs and CMs.

Key words: bone marrow mesenchymal stem cells (MSCs), cardiomyocytes (CMs), co-culture microenvironment, metabonomics, myocardium-like differentiation

CLC Number:

O658

HE Linjing, DENG Fuxue, HU Yunfeng, ZHU Weiwen, WU Jiayan, CHANG Jing. Feasibility of cardiac differentiation of bone marrow mesenchymal stem cells induced by co-culture indirectly with cardiomyocytes in several dimensions[J]. Chinese Journal of Chromatography, 2016, 34(4): 414-421.

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