Culturing Schwann cells by explants and modified enzyme digestion technique

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Author:
WANG Hui(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
LUO Lun(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
LI Wen-sheng(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
LIANG Chao-feng(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
HE Hai-yong(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
GUO Ying(Department of Neurosurgery,Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)
Journal Title:
CHINESE JOURNAL OF NEUROMEDICINE
Issue:
Volume 10, Issue 05, 2011
DOI:
10.3760/cma.j.issn.1671-8925.2011.05.008
Key Word:
Schwann cell;Cell culture technology;Enzyme digestion;Explant

Abstract: Objective To introduce an efficient method for culturing and purifying Schwann cells (SCs) in vitro. Methods Sciatic nerves were harvested from neonatal SD rats and the epineuria were removed. Single enzyme digestion combined with explans and double enzyme digestion were employed, respectively, to digest the nerve tissues following trituration. The proliferation of SCs and degree of purification were evaluated by viable count method and the combination of S-100 labeled monoclonal antibody and SCs. Results Proximally 3.5 × 106 cells were harvested with 96% survival rate and a purity of Schwann cells over 94% by single enzyme digestion combined with explans, however only 3.0×106 cells were gained with the purity being 90% and survival rate being 92% by double enzyme digestion. Conclusion This method yields large amount of viable Schwann cells with high purity and survival rate.

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