Chinese Journal of Tissue Engineering Research ›› 2023, Vol. 27 ›› Issue (19): 3017-3022.doi: 10.12307/2023.667

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Obtaining microglia-like cells after mouse corneal epithelial cell induction

Xu Huifang, Shan Ping, Li Lijuan   

  1. Department of Neurology, Wuhan No. 1 Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430000, Hubei Province, China
  • Received:2022-08-01 Accepted:2022-09-19 Online:2023-07-08 Published:2022-11-28
  • Contact: Xu Huifang, PhD, Attending physician, Department of Neurology, Wuhan No. 1 Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430000, Hubei Province, China Shan Ping, PhD, Associate Chief physician, Department of Neurology, Wuhan No. 1 Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430000, Hubei Province, China
  • About author:Xu Huifang, PhD, Attending physician, Department of Neurology, Wuhan No. 1 Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430000, Hubei Province, China
  • Supported by:
    National Natural Science Foundation of China, No. 81703870 (to SP); a grant from Wuhan Municipal Health Commission, No. WX16B01 (to XHF); the General Program of Wuhan Science and Technology Bureau, No. 2015061701011618 (to XHF)

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Abstract: BACKGROUND: Microglial cells are the predominant immune cells of the central nervous system and are important for the study of the development and treatment of many diseases. However, the methods for obtaining individualized cells in vitro are still lacking.  
OBJECTIVE: To establish a new microglia induction culture system to induce mouse corneal epithelial cells into microglia-like cells.
METHODS: Corneal epithelial tissues of 2-week-old mice were lysed to obtain epithelial cells, and modified media containing interleukin-34 (100 ng/mL), colony stimulating factor 1 (5 ng/mL), transforming growth factor β (1 ng/mL), interleukin-6 (1 ng/mL), and recombinant vascular endothelial growth factor (1 ng/mL) were successively added. Treatment lasted up to 16 days. Cell properties were evaluated after induction by transcriptional level, protein expression level, subcellular localization, flow cytometry and morphological observation.  
RESULTS AND CONCLUSION: After induction, microglia-like cells which express characteristic molecules of TMEM119, CXCL11, CD11b and CD68 were obtained. Morphological observation, flow cytometry and fluorescence analysis confirmed that the expression products of TMEM119, CXCL11, CD11b and CD68 microglia had high abundance, which was very similar to microglia.

Key words: mouse corneal epithelial cell, microglial cell, induced differentiation, TMEM119, CD68, CXCL11, CD11b

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