Direct reprogramming of human fibroblasts into sweat gland-like cells

Abstract

These authors contributed equally to this work.Keywords: fibroblast, Lef-1, NF-kB, reprogramming, regeneration, sweat glands Abbreviations: CEA, carcinoembryonic antigen; CK7, cytokeratin 7; CK14, cytokeratin 14; CK19, cytokeratin 19; DMEM, Dulbecco's modified Eagle's medium; FBS, fetal bovine serum; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Lef-1, lymphoid enhancer-binding factor 1; min, minutes; NF-kB, nuclear factor kappa-light-chain-enhancer of activated B cells; Shh, sonic hedgehogThe skin of patients with an extensive deep burn injury is repaired by a process that leaves a hypertrophic scar without sweat glands and therefore loses the function of perspiration. The aim of this study was to identify whether the key factors related to sweat gland development could directly reprogram fibroblasts into sweat gland-like cells. After introducing the NF-kB and Lef-1 genes into fibroblasts, we found that stably transfected fibroblasts expressed specific markers of sweat glands, including CEA, CK7, CK14 and CK19, both at the protein and mRNA levels. The immunofluorescence staining also showed positive expression of CEA, CK7, CK14 and CK19 in induced fibroblasts, but there were no positive cells in the control groups. The expression of Shh and Cyclin D1, downstream genes of NF-kB and Lef-1, were also significantly increased during regeneration. The induced fibroblasts were implanted into an animal model. Twenty days later, iodine-starch perspiration tests showed that 7 out of the 10 cell-treated paws were positive for perspiration, with a distinctive black point-like area appearing in the center of the paw. Contralateral paws tested negative. Histological examination of skin biopsies from experimental and control paws revealed that sweat glands were fully reconstructed in the test paws, with integral, secretory and ductal portions, but were not present in the control paws. This is the first report of successful reprogramming of fibroblasts into sweat gland-like cells, which will provide a new cell source for sweat gland regeneration in patients with extensive deep burns.

The evolution of eccrine sweat gland research towards developing a model for human sweat gland function

“…The initial report of reprogrammed human fibroblasts, transplanted into the paws of mice with full thickness scald injuries becoming functional sweat gland cells, is an interesting development of the findings that sweat gland cells contribute to wound healing. However, these results require further investigation, as it is not yet clear whether the detected sweat gland cells were derived from incomplete removal of glands from the host or truly derived from the transplanted fibroblasts.…”

Regenerative and reparative effects of human chorion-derived stem cell conditioned medium on photo-aged epidermal cells

“…Many methods have been used for skin repair, such as stem cell injection and somatic cells reprogramming for tissue regeneration. 26 However, efficacious and safe treatments to repair photo-injured skin need further research. In this study, CDSC-CNM has shown efficient regenerative and reparative effects on UVB-irradiated keratinocytes.…”

Bioactive Molecules for Skin Repair and Regeneration: Progress and Perspectives

“…A recent research suggested that the human keratinocyte growth factor played a pivotal role in promoting hucMSC differentiation to sweat gland-like cells [95]. The first transgenic report transformed the key factors NF-kb and Lef-1 that were related to sweat gland development and directly reprogrammed fibroblasts into sweat gland-like cells [96]. However, no study reported on how to use bioactive molecules alone to obtain the SGCs, which needed further exploration.…”