Fig. 3

Conversion of HDF into iSGC with small molecules. a Scheme of reprogramming strategy procedure. HDF was transfected with EDA and plated in fibroblasts medium, then cells were transferred into SGM for 15–20 d, and eventually HDF-EDA in SGM were further transferred into mSGM for 6–8 d. b qPCR analysis of transcriptional expression of LEF1, TGFβ-R1, BMPR1A, β₂-AR and RARα in HDF and pSGC; The genes showing significant differences were observed. c Phase contrast images showing the morphological changes of iSGC in mSGM on day 3 and 6. Scale bar = 200 μm. Illustrations, higher magnification of the boxed areas. d qPCR analysis of transcriptional expression of CK5, CK10, CK18, AQP5 and CEA in HDF and iSGC after 6—8 d of induction. e, f Flow cytometry quantification of CK5 positive cells and CK18 positive cells of iSGC. n = 3. Data were expressed as mean ± SD and analyzed by two-tailed t-tests, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001. EDA ectodermal dysplasia antigen, SGM sweat gland culture medium, EGF epidermal growth factor, bFGF basic fibroblast growth factor, RCIRB (Repsox, CHIR99021, Isoproterenol, Retinoic acid, BMP4), HDF human dermal fibroblasts, pSGC primary sweat gland cell, LEF1 lymphoid enhancer-binding factor 1, TGFβ-R1 TGF-β type 1 receptor, BMPR1A bone morphogenetic protein receptor 1A, β₂-AR β₂-adrenergic receptor, RARα retinoic acid receptor α, iSGC induced sweat gland-like cell, mSGM modified sweat gland culture medium, CK5 cytokeratin 5, CK10 cytokeratin 10, CK18 cytokeratin 18, AQP5 aquaporin 5, CEA carcino-embryonic antigen