Combination of 5‐azaytidine and hanging drop culture convert fat cell into cardiac cell,Biotechnology and Applied Biochemistry

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Combination of 5‐azaytidine and hanging drop culture convert fat cell into cardiac cell
Biotechnology and Applied Biochemistry ( IF 3.2 ) Pub Date : 2020-02-06 , DOI: 10.1002/bab.1897
Zahra Bagheri-Hosseinabadi _{1,

2} , Fatemeh Seyedi ₃ , Hamid Reza Mollaei ₄ , Mojgan Moshrefi ₅ , Alexander Seifalian ₆

Affiliation

One of the promising approaches for the treatment of cardiac disease is stem cell therapy. In this study, we compared the cardiomyogenic differentiation rate, from human adipose‐derived stem cells (hADSCs) in a three‐dimensional (3D) hanging drop (HD) spheroid culture system, versus a two‐dimensional (2D) culture condition at different concentrations of 5‐azacytidine (5‐Aza). 5‐Azaytidine (5‐Aza) is a pyrimidine nucleoside analogue of cytidine that initiates cell differentiation programs through DNA demethylation. The hADSCs were isolated and cultured both in 2D and 3D HD conditions, with either 10 or 50 μM concentrations of 5‐Aza. Then DNA content, gene expression, and protein content were analyzed. 3D HD culture resulted in a higher percentage of cells in G0/G1 and S phase in the cell division cycle, whereas 2D culture led to a greater percentage of cells in the G2/M phase. A significantly higher gene expression rate of HAND1, HAND2, cTnI, Cx43, βMHC, GATA4, NKX2.5, and MLC2V was observed in HD treated with 50 μM 5‐Aza. This was confirmed by immunocytochemistry. These findings suggest that 50 μM concentration of 5‐Aza can induce hADSCs to differentiate into cardiomyocytes. The differentiation rate was significantly higher when accompanied by the 3D HD culture system. This work provides a new culture system for cell differentiation for cardiovascular tissue engineering.

中文翻译：

5-氮杂胞苷与悬滴培养的结合将脂肪细胞转化为心脏细胞

干细胞疗法是治疗心脏病的有前途的方法之一。在这项研究中，我们比较了在3维（3D）悬滴（HD）球状培养系统中人脂肪干细胞（hADSCs）与不同条件下二维（2D）培养条件下的心肌分化率浓度的5-氮杂胞苷（5-氮杂）。5-氮杂胞苷（5-Aza）是胞嘧啶的嘧啶核苷类似物，可通过DNA脱甲基作用启动细胞分化程序。分离了hADSC，并在2D和3D HD条件下用10或50μM浓度的5-氮杂进行培养。然后分析DNA含量，基因表达和蛋白质含量。3D HD培养导致细胞分裂周期中G0 / G1和S期的细胞百分比更高，而2D培养导致G2 / M期的细胞百分比更高。在用50μM5-Aza处理的HD中，观察到HAND1，HAND2，cTnI，Cx43，βMHC，GATA4，NKX2.5和MLC2V的基因表达率明显更高。免疫细胞化学证实了这一点。这些发现表明，浓度为50μM的5-氮杂可以诱导hADSCs分化为心肌细胞。当与3D HD培养系统一起使用时，分化率明显更高。这项工作为心血管组织工程的细胞分化提供了一种新的培养系统。这些发现表明，浓度为50μM的5-氮杂可以诱导hADSCs分化为心肌细胞。当与3D HD培养系统配合使用时，分化率明显更高。这项工作为心血管组织工程的细胞分化提供了一种新的培养系统。这些发现表明，浓度为50μM的5-氮杂可以诱导hADSCs分化为心肌细胞。当与3D HD培养系统一起使用时，分化率明显更高。这项工作为心血管组织工程的细胞分化提供了一种新的培养系统。

更新日期：2020-02-06

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