Isolation of human mesenchymal stem cells from the skin and their neurogenic differentiation in vitro
변준호, 강은주, 박성철, 강동호, 최문정, 노규진, 박봉욱,
소속 상세정보
변준호 ( Byun Jun-Ho ) - Gyeongsang National University School of Medicine Department of Oral and Maxillofacial Surgery
강은주 ( Kang Eun-Ju ) - Gyeongsang National University College of Veterinary Medicine OBS/Theriogenology and Biotechnology
박성철 ( Park Seong-Cheol ) - Gyeongsang National University School of Medicine Department of Oral and Maxillofacial Surgery
강동호 ( Kang Dong-Ho ) - Gyeongsang National University School of Medicine Department of Neurosurgery
최문정 ( Choi Mun-Jeong ) - Gyeongsang National University School of Medicine Department of Oral and Maxillofacial Surgery
노규진 ( Rho Gyu-Jin ) - Gyeongsang National University College of Veterinary Medicine OBS/Theriogenology and Biotechnology
박봉욱 ( Park Bong-Wook ) - Gyeongsang National University School of Medicine Department of Oral and Maxillofacial Surgery
KMID : 0355620120380060343
Abstract
Objective: This aim of this study was to effectively isolate mesenchymal stem cells (hSMSCs) from human submandibular skin tissues (termed hSMSCs) and evaluate their characteristics. These hSMSCs were then chemically induced to the neuronal lineage and analyzed for their neurogenic characteristics in vitro.
Materials and Methods: Submandibular skin tissues were harvested from four adult patients and cultured in stem cell media. Isolated hSMSCs were evaluated for their multipotency and other stem cell characteristics. These cells were differentiated into neuronal cells with a chemical induction protocol. During the neuronal induction of hSMSCs, morphological changes and the expression of neuron-specific proteins (by fluorescence-activated cell sorting [FACS]) were evaluated.
Results: The hSMSCs showed plate-adherence, fibroblast-like growth, expression of the stem-cell transcription factors Oct 4 and Nanog, and positive staining for mesenchymal stem cell (MSC) marker proteins (CD29, CD44, CD90, CD105, and vimentin) and a neural precursor marker (nestin). Moreover, the hSMSCs in this study were successfully differentiated into multiple mesenchymal lineages, including osteocytes, adipocytes, and chondrocytes. Neuron-like cell morphology and various neural markers were highly visible six hours after the neuronal induction of hSMSCs, but their neuron-like characteristics disappeared over time (24-48 hrs). Interestingly, when the chemical induction medium was changed to Dulbecco’s Modified Eagle Medium (DMEM) supplemented with fetal bovine serum (FBS), the differentiated cells returned to their hSMSC morphology, and their cell number increased. These results indicate that chemically induced neuron-like cells should not be considered true nerve cells.
Conclusion: Isolated hSMSCs have MSC characteristics and express a neural precursor marker, suggesting that human skin is a source of stem cells. However, the in vitro chemical neuronal induction of hSMSC does not produce long-lasting nerve cells and more studies are required before their use in nerve-tissue transplants.
키워드
Skin; Mesenchymal stem cell; In vitro neuronal differentiation
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