Efficient Schwann cell purification by differential cell detachment using multiplex collagenase treatment.
Schwann cell purification is usually difficult due to the contamination of fibroblasts, which often become a predominant cell type in Schwann cell culture in vitro. We have developed a novel and efficient method to enrich Schwann cells by differential detachment of two types of cells. In the culture, cells were treated with a multiplex collagenase and the Schwann cells were found to detach faster than fibroblasts and thus Schwann cells could be easily isolated. Within 5 days, Schwann cell purity could reach above 99%, which was confirmed by immunostaining characterization and flow cytometric analysis. In addition, this efficient method can reach a high cell yield after two rounds of differential detachment procedures and does not require antimitotic treatment or special equipment as often needed by other reported methods.
Jin
Yu-Qing
YQ
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Tissue Engineering Laboratory, Shanghai, China.
Liu
Wei
W
Hong
Tan-Hui
TH
Cao
Yilin
Y
D000818:Animals; D002452:Cell Count; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D020800:Receptor, Nerve Growth Factor / Q000502:physiology; D012583:Schwann Cells / Q000187:drug effects / Q000502:physiology*
D020800:Receptor, Nerve Growth Factor; D017364:Collagenases