Journal of Shandong University (Health Sciences) ›› 2019, Vol. 57 ›› Issue (11): 1-8.doi: 10.6040/j.issn.1671-7554.0.2019.813

   

Comparison of DNA methylation levels between three-dimensional and two-dimensional cultured umbilical cord mesenchymal stem cells

HUANG Jinxian1, LI Dong2, LI Cong1, SHI Qing2, JU Xiuli1   

  1. 1. Department of Pediatrics, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
    2. Stem Cell and Regenerative Medicine Research Center, Shandong University, Jinan 250012, Shandong, China
  • Published:2022-09-27

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Abstract: Objective To explore the effects of three-dimensional(3D)culture on the whole genome DNA methylation profile of umbilical cord mesenchymal stem cells(UC-MSCs). Methods UC-MSCs were isolated from umbilical cords using tissue adherent culture. Their phenotypes were identified with flow cytometry. Then UC-MSCs were cultured in 3D polyvinyl fluoride nested scaffolds and 2D conventional culture flask, respectively. Acridine orange was used to observe UC-MSCs on the scaffold. Methylated DNA immunoprecipitation(MeDIP-Seq)method was used to detect the whole genome DNA methylation of UC-MSCs. Results UC-MSCs were successfully isolated from umbilical cords and able to grow on the 3D scaffold. The genomic DNA methylation level of UC-MSCs increased after 3D culture. Gene ontology analysis showed that the up-regulated differential methylation genes were involved in protein metabolism, membrane-related organelle composition, and enzyme activity regulation. Down-regulated differential methylation genes were involved in vesicle trafficking and cGMP metabolic processes regulating. According to pathway analyses, differential methylation genes were involved in RNA transport, neuroactive ligand-receptor interaction, cellular communication, and 山 东 大 学 学 报 (医 学 版)57卷11期 -黄金献,等.三维和二维培养的脐带间充质干细胞DNA甲基化水平比较 \=-vesicle secretion-related pathways. Conclusion 3D culture changes the DNA methylation profile of UC-MSCs. The up-regulated methylation levels will alter the function of cell migration and proliferation. 3D culture reduces the methylation level of immune regulation and secretion of vesicle-related genes in UC-MSCs.

Key words: Mesenchymal stromal cells, Three-dimensional culture, DNA methylation, methylated DNA immunoprecipitation sequencing

CLC Number: 

  • R574
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