Enrichment of epidermal stem cells of rats by Vario magnetic activated cell sorting system |
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Authors: | Wei Chen Wei-wei Zhang Chunying Shi Xiaohua Lian Shanghong Yi Tian Yang |
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Affiliation: | 1. Department of Urology, Xinqiao Hospital, The Third Military Medical University, Chongqing, 400037, China 2. Department of Nephrology, Daping Hospital, The Third Military Medical University, Chongqing, 400042, China 3. Department of Cell Biology, The Third Military Medical University, Chongqing, 400038, China
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Abstract: | Epidermal stem cells (ESCs) play an important role in skin homeostasis, wound repair, and tumorigensis which have great potential in scientific research and clinical application. So, the efficient isolation of these infrequent stem cells is very important for researchers to solve the problem of low purity and insufficient quantity of stem cells in vitro. The aim of this study was to investigate a method for the enrichment of ESCs by magnetic activated cell sorting system. The isolation strategy was CD71 depletion followed by α6-integrin positive selection. The percentage of α6briCD71dim cells in isolated cells was 94.59%. Transmission electron microscopy results revealed that α6bri CD71dim cells exhibited some typical characteristics like progenitor cells, such as big nucleus, obvious nucleolus, large nuclear–cytoplasm ratio, and few organelles in cytoplasm. When cultured in vitro, the α6briCD71dim cells had greater proliferating potential and higher colony-forming ability, and high levels of epidermal stem cell markers were expressed in our positive cells. ESCs have been successfully isolated from neonatal epidermis using Vario MACS and cultured in vitro. This isolation method is simple, fast, and inexpensive, providing an important tool for tissue engineering and cell transplantation studies. |
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