Isolation,culture and characterization of neural crest cells from lung tissue of mT/mG;Wnt1-Cre mice
10.16352/j.issn.1001-6325.2024.11.1510
- VernacularTitle:mT/mG;Wnt1-Cre小鼠肺组织神经嵴细胞的分离培养与特征鉴定
- Author:
Xiaowen DONG
1
;
Yongxin LI
;
Xiaoxue GONG
;
Lingfang FENG
;
Junfei CHEN
;
Jiahui YAO
;
Jianlin LOU
Author Information
1. 杭州医学院 公共卫生学院,浙江 杭州 310013
- Keywords:
neural crest cells;
primary culture;
flow cytometry;
SRY-box transcription factor 10(Sox10)
- From:
Basic & Clinical Medicine
2024;44(11):1510-1515
- CountryChina
- Language:Chinese
-
Abstract:
Objective To isolate and culture neural crest cells(NCCs)from lung tissue of mice and to identify the characteristics of the cells in order to provide a new cell model for studying lung injury and injure repair.Methods The mT/mG dual-fluorescence reporter mice and Wnt1-Cre transgenic mice were hybridized,and mT/mG;Wnt1-Cre transgenic mice were screened to obtain enhanced green fluorescent protein(EGFP)permanently labeled NCCs.Cell suspension of mouse lung tissue was prepared by enzymolysis.EGFP+cells(namely NCCs)were har-vested by flow cytometry.Primary culture was performed with DMEM/F12 culture medium optimized in the labora-tory,NCCs was characterized by immunofluorescence microscopy.Then NCCs differentiation was directed by mouse bone marrow mesenchymal stem cells osteogenic induction.Results The mT/mG of EGFP permanently labeled NCCs was successfully obtained by hybridization and high-purity NCCs were isolated from Wnt1-Cre transgenic mice lung tissue.They can be cultured in vitro and with spindle morphology which was,similar to fibroblast adherent proliferation.NCCs expressed the neural crest stem cell marker Sox10 and induced to differentiate into osteoblasts.Conclusions NCCs isolated and cultured from lung tissue of mT/mG;Wnt1-Cre transgenic mice show stable prolif-eration and have the characteristics of neural crest stem cells,which may function as a potential cell model for re-search on lung tissue injury and the mechanism of repair.
