Combined use of adipose-derived stem cells and electrospun membrane for preparing sandwich-like cardiac cell sheets

doi:10.3969/j.issn.2095-4344.0015

Abstract

Abstract:

BACKGROUND: Increasing studies have shown that cell-sheet engineering is a more promising method for the treatment of myocardial infarction.

OBJECTIVE: To construct sandwich-like myocardial cell sheets with electrospun chitosan-collagen membrane as support membrane and with adipose-derived stem cells as seed cells.

METHODS: In this study, we prepared electrospun chitosan-collagen membranes at different mass ratio (7:3, 5:5) as a support membrane. Adipose-derived stem cells at passage 4 that were cultured in a thermo-sensitive petri dish for 3 days were seeded onto the two kinds of electrospun membranes at 20 ℃ for 15 minutes until the cell sheets rolled up. Then, the electrospun membranes and cell sheets with the cell layers facing up were placed together onto another thermo-sensitive petri dish in which adipose-derived stem cells were confluent completely. After 30 minutes of culture, the cell sheets were cultured in myocardial cell medium. Two weeks later, cell morphology was observed using multi-photon microscopy. Immunocytochemistry, western blot and flow cytometry were used to detect the expression of TnI and Cx43 in the myocardium. Real-time quantitative PCR was used to detect cardiomyocyte-specific gene expression.

RESULTS AND CONCLUSION: From the results of scanning electronic microscopy (SEM) and propidium iodide (PI) staining, we could found that ADSCs grew easily on the chitosan-collagen (5:5) membrane, and there were more dead cells on the chitosan-collagen (7:3) membrane than on the chitosan-collagen (5:5) membrane. After 2 weeks of differentiation with the myocardial cell medium, higher troponin I and Cx43 protein expressions were observed on the chitosan-collagen (5:5) membrane (P < 0.05). Moreover, the mRNA levels of α-skA, β-MHC, TnI, Cx43, ANP, GATA-4 and Nkx2.5 were higher in the chitosan-collagen (5:5) membrane group than the chitosan-collagen (7:3) membrane group. All these results indicate that the chitosan-collagen (5:5) membrane is better for the growth and differentiation of adipose-derived stem cells, as well as for sandwich-like cell-sheet construction.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Chitosan, Stem Cells, Myocardial Infarction, Tissue Engineering

CLC Number:

R318

Wu Zhang-song, Zhu Hong-xia, Luo Zhi-qiang, Pan Wan-chun, Yang Fang, Lin Xin-yan, Zhu Yan-xia.

Combined use of adipose-derived stem cells and electrospun membrane for preparing sandwich-like cardiac cell sheets

[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(2): 254-260.

Figures/Tables 9

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