Expression of autoimmune regulator during differentiation of mouse embryonic stem cells into thymic epithelial progenitor cells

doi:10.3969/j.issn.2095-4344.2111

Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (25): 3994-3999.doi: 10.3969/j.issn.2095-4344.2111

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Expression of autoimmune regulator during differentiation of mouse embryonic stem cells into thymic epithelial progenitor cells

Ma Jingyi, Yang Wenjiang, Li Yuandi, Huang Youjiao, Gao Jie, Li Hong, Hu Rong, Su Min

Department of Histology and Embryology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou Province, China

Received:2020-01-19 Revised:2020-01-20 Accepted:2020-03-05 Online:2020-09-08 Published:2020-08-24
Contact: Corresponding author: Hu Rong, MD, Professor, Department of Histology and Embryology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou Province, China Corresponding author: Su Min, MD, Professor, Department of Histology and Embryology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou Province, China
About author:Ma Jingyi, Master, Department of Histology and Embryology, College of Basic Medical Sciences, Guizhou Medical University, Guiyang 550025, Guizhou Province, China
Supported by:
the National Natural Science Foundation of China, No. 81660033 ; the Science and Technology Supporting Program of Guizhou Province, No. [2017]2970 ; Program for Science and Technology Excellent Talents in Universities of Guizhou Province, No. [2017]071 ; the Science and Technology Cooperation Project of Guizhou Province, No. [2016]7363

Abstract

Abstract:

BACKGROUND: Autoimmune diseases are a class of diseases that cause a strong immune response to the continuous lack of self-tissue-specific antigens in the thymus. Hypothyroidism and unstable expression of tissue-specific antigens in the thymus can limit the therapeutic effect. The thymus is mainly composed of thymic epithelial cells, but the limited number of mature thymic epithelial cells and thymic epithelial progenitor cells in the thymus has greatly limited related research.

OBJECTIVE: To detect the expression of autoimmune regulator (AIRE) when mouse embryonic stem cells were transformed into thymic epithelial progenitor cells.

METHODS: A two-step differentiation method was used to induce the differentiation of mouse embryonic stem cells into endoderm and then into thymic epithelial progenitor cells. The cells were collected at 0, 3, and 13 days of induced differentiation. Immunofluorescence, flow cytometry, western blot and real-time PCR were used to detect the expression of cell-associated genes and proteins.

RESULTS AND CONCLUSION: Positive expression of OCT4 and SSEA1 was detected by immunofluorescence at 0 day of induction. The double positive expression of SOX17 and FoxA2 was measured by immunofluorescence at 3 days of induction. The positive expression of EpCAM, K5 and K8 were analyzed by flow cytometry at 13 days of induction. During the directional differentiation of mouse embryonic stem cells, real-time PCR indicated that the expression of PAX1, PAX9, FOXN1 and PLET1 showed an increasing trend. The expression of AIRE gene increased significantly at 0, 3, and 13 days of induction. At the same time, the expression of INS2 gene and GAD67 gene also increased. Western blot assay showed that the expression of AIRE protein gradually decreased at 0, 3, and 13 days of induction; however, insulin protein and GAD67 protein were not detected. Overall findings indicate that mouse embryonic stem cells can successfully differentiate into thymic epithelial progenitor cells with highly expressed AIRE gene, which promotes the expression of INS2 and GAD67 genes, and provides an evaluation basis for cell transplantation in the treatment of autoimmune diseases.

Key words:

embryonic stem cells, autoimmune regulator, thymic epithelial progenitor cells, tissue-specific antigens

CLC Number:

Ma Jingyi, Yang Wenjiang, Li Yuandi, Huang Youjiao, Gao Jie, Li Hong, Hu Rong, Su Min. Expression of autoimmune regulator during differentiation of mouse embryonic stem cells into thymic epithelial progenitor cells [J]. Chinese Journal of Tissue Engineering Research, 2020, 24(25): 3994-3999.

Figures/Tables 8

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Expression of autoimmune regulator during differentiation of mouse embryonic stem cells into thymic epithelial progenitor cells

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