ABSTRACT
Cell therapy has shown its power to promote diabetic chronic wound healing. However, problems of scar formation and loss of appendages have not yet been solved. Our study aims to explore the potential of using embryonic skin cells (ESkCs) to repair diabetic wounds. Circular wound was created on the back of the diabetic mice, and ESkCs stained with CM-DIL were transplanted into the wound. Wound area was recorded at the day 4, 7, 11, and 14 after transplantation. The tissue samples were obtained at week 1, 2, and 3, and the tissue sections were stained by transforming growth factor ß1 (TGF-ß1), TGF-ß3, vascular endothelial growth factor (VEGF), and CD31. The new skin formed on the wound of the diabetic mice with ESkC treatment at week 1 but not on the wounds of the non-treatment group. The histological scores of diabetic group with ESkC treatment were significantly better than the non-treatment group (P < 0.05). The fluorescence examination of CM-DIL and CD31 staining indicated that the ESkCs participated in the tissue regeneration, hair follicles formation, and angiogenesis. The expression of TGF-ß1 and VEGF in ESkC-treated groups was noticeable in week 1 but disappeared in week 2. TGF-ß3 was not expressed at week 1 but expressed markedly around hair follicles in week 2 in ESkC-treated groups. Our study demonstrated that ESkCs are capable of developing new skin with appendage restoration to repair the diabetic wounds.
Subject(s)
Cell Transplantation/methods , Diabetes Complications , Wounds and Injuries/therapy , Animals , Disease Models, Animal , Histocytochemistry , Immunohistochemistry , Male , Mice, Inbred C57BL , Microscopy , Skin/anatomy & histology , Skin Physiological Phenomena , Wound HealingABSTRACT
Human embryonic stem cells (ESCs) can differentiate into endothelial cells in response to stimuli from extracellular cytokines. Transforming growth factor (TGF)-ß1 signaling is involved in stem cell renewal and vascular development. Previously, human ESCs were isolated from inner cell mass and a stable ESC line was developed. In the present study, the effects of extracellular TGF-ß1 were investigated on human ESC-derived embryoid bodies (EB) in suspension. The structures of the EBs were analyzed with light and electron microscopy, while the cellular composition of the EBs was examined via the expression levels of specific markers. Vascular-like tubular structures and cardiomyocyte-like beating cells were observed in the EBs at day 3 and 8, respectively. The frequencies of vascular-like structures and beating cells in the TGF-ß1 treated group were significantly higher compared with the control group (84.31 vs. 12.77%; P<0.001; 37.25 vs. 8.51%; P<0.001, respectively). Electron microscopy revealed the presence of lumens and gap junctions in the sections of the tubular structures. Semiquantitative polymerase chain reaction revealed elevated expression levels of CD31 and fetal liver kinase-1 in EBs cultured with TGF-ß1. In addition, extensive staining of von Willebrand factor was observed in the vascular-like structures of TGF-ß1-treated EBs. Therefore, the results of the present study may aid the understanding of the underlying mechanisms of human ESC differentiation and improve the methods of propagating specific cell types for the clinical therapy of cardiovascular diseases.
