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[Sorting and identification of human immature Langerhans cells]
Jianbin Wang1, Lichun Wang1, Lei Liu1
1Yunnan Key Laboratory of Vaccine Research and Development on Severe Infectious Disease; Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118, China.
Abstract:
Objective To isolate the immature Langerhans cells(LCs) of human epidermis and to identify the phenotype based on their surface marker. Methods After treatment of piecemeal tissues of human foreskin with dispase I digesting overnight, the separated epidermis was further digested by collagenaseIat 37DegreesCelsius in a shaker for 2 hours to produce a suspension of dispersive epithelium, in which, LCs population of CD3-CD14-CD1a+CD45+ was harvested in flow cytometry influx with certain antibodies and identified further by immunofluorescence assay of anti-CD207 antibody as while the vitality of these cells were observed with trypan blue staining. Results The population containing 91.31% of immature LCs was sorted and harvested, which showed 93.16% of survival rate. Conclusion Technical procedure of sorting immature LCs from human foreskin was established and ascertained being effective.
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