Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method.
Objective To develop an improved method for culturing Schwann cells(SCs) by using both enzyme digestion and explants-culture approaches and compared with traditional explants-culture method and general hemi-explants-culture method. Methods Bilaterally sciatic nerves and brachial plexus nerves were dissected from 3 to 5-day-old neonatal SD rats and explants-culture method,general hemi-explants-culture method,and improved enzyme digestion combined with explants-culture method were adopted to culture SCs,respectively. SCs were digested and passaged after 7 days in culture and counted under the microscope. The purity of SCs was identified by S-100 immunofluorescence staining. Results The SCs of improved method group grew fastest and the total number of cells obtained was(1.85±0.13)×10(6);the SCs of the hemi-explants-culture method group grew slower than the improved method group and the total number of cells obtained was (1.10±0.10)×10(6);the SCs of the explants-culture method group grew slowest and the total number of cells obtained was (0.77±0.03)×10(6).The total number of cells obtained showed significant difference among the three groups(P<0.01). Immunofluorescence staining showed that the SCs purity was (95.73±1.51)% in the improved method group,(84.66±2.68)% in the hemi-explants-culture method group,and (74.50±4.23)% in the explants-culture method group(P<0.01). Conclusion The improved enzyme digestion combined with explants-culture method can obtain sufficient amount of high-purity SCs in a short time and thus may be applied in further research on peripheral nerve regeneration.
Department of Traditional Chinese Medicine,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.
Di
D
Liu
Department of Traditional Chinese Medicine,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.
Xiao-Chun
XC
Liang
Department of Cell Resource Center,Institute of Basic Medical Science,CAMS and PUMC,Beijing 100005,China.
Hong
H
Zhang
D000818:Animals; D000831:Animals, Newborn; D001917:Brachial Plexus / Q000166:cytology; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004798:Enzymes; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000166:cytology*; D012584:Sciatic Nerve / Q000166:cytology
D004798:Enzymes