2025, Vol. 29 ›› Issue (36): 7790-7796

Transcription factor NKX2.1 promotes differentiation of induced pluripotent stem cells into lung stem cells

Deng Li, Liu Yang, Wang Hui, Yang Qiu, Dong Mingqing   

Abstract: BACKGROUND: Enhancing the differentiation of induced pluripotent stem cells into lung stem cells is crucial for repairing lung injuries. NKX2.1 is the earliest marker of lung epithelial differentiation and plays a significant regulatory role in lung development. However, the impact of its expression on the differentiation of induced pluripotent stem cells into lung stem cells remains inadequately understood.
OBJECTIVE: To investigate the effect of NKX2.1 on the differentiation of induced pluripotent stem cells into lung stem cells.
METHODS: Induced pluripotent stem cells were cultured in vitro. The expression of specific pluripotent stem cell genes was assessed using real-time fluorescence quantitative PCR. NKX2.1 was overexpressed in induced pluripotent stem cells, which were then induced to differentiate into lung stem cells. The expression of FoxA2, SOX9, and P63 was determined via quantitative PCR and immunofluorescence on day 7 of induction of differentiation. The expression of the alveolar marker SPB and SPC was evaluated through immunofluorescence staining on day 7 of induction of differentiation.
RESULTS AND CONCLUSION: (1) Induced pluripotent stem cells in vitro were tightly packed and showed typical clonoid growth and significantly expressed stem cell-specific genes OCT-4, SOX2, and NANOG. (2) Compared with the non-transfected control group, the expression of NKX2.1 in human induced pluripotent stem cells was significantly increased in the NKX2.1 overexpression group (P < 0.000 1). (3) Seven days after induction of differentiation, compared with the non-transfected control group, the expression of lung stem cell-related markers FoxA2, SOX9, and P63 was significantly increased in the NKX2.1 overexpression group (P < 0.000 1). (4) Thirteen days after induction of differentiation, compared with the non-transfected control group, the fluorescence intensity of alveolar cell marker molecules SPB and SPC increased significantly in the overexpression NKX2.1 group. The results show that NKX2.1 can promote the differentiation of induced pluripotent stem cells into lung stem cells.  

Key words: induced pluripotent stem cell, lung stem cell, NKX2.1, definitive endoderm, foregut endoderm, lung injury, engineered stem cell