Abstract: The inaccessibility of the human fetus has led to the development of in vitro methods for generating human embryonic neural crest cells (NCCs), providing valuable insights into their cellular and molecular characteristics. This approach enables the modeling of neural crest defects for mechanistic studies and identification of therapeutic targets. While previous efforts have involved invasive procedures to directly reprogram human blood cells and fibroblasts into NCCs, this study explores a non-invasive approach using urine cells for reprogramming into NC-like cells by lentiviral-mediated overexpression of two different polycistronic cassettes, BRN2-KLF4-SOX2-ZIC3 (BKS2Z) and BRN2-KLF4-SOX17-ZIC3 (BKS17Z). The results show that BKS17Z is more effective than BKS2Z in generating NC-like cells from urine cells. Moreover, the addition of the small molecule DAPT enhances the reprogramming efficiency of BKS17Z compared to direct differentiation of induced pluripotent stem cells into NC-like cells. The reprogrammed NC-like cells exhibit expression of cranial neural crest markers, migratory capacity and the ability to differentiate into neural crest derivatives such as chondrocytes. Altogether, we have established a non-invasive strategy to reprogram urine cells into cranial NC-like cells, paving the way for modeling craniofacial disorders.
Funding Source: Research Grants Council and University Council of Hong Kong (GRF_17102420 & GRF_17114619)
