Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method.
10.3881/j.issn.1000-503X.2016.04.004
- Author:
Di LIU
1
;
Xiao-Chun LIANG
1
;
Hong ZHANG
2
Author Information
1. Department of Traditional Chinese Medicine,PUMC Hospital,CAMS and PUMC,Beijing 100730,China.
2. Department of Cell Resource Center,Institute of Basic Medical Science,CAMS and PUMC,Beijing 100005,China.
- Publication Type:Journal Article
- MeSH:
Animals;
Animals, Newborn;
Brachial Plexus;
cytology;
Cell Culture Techniques;
Cell Separation;
methods;
Cells, Cultured;
Enzymes;
Rats;
Rats, Sprague-Dawley;
Schwann Cells;
cytology;
Sciatic Nerve;
cytology
- From:
Acta Academiae Medicinae Sinicae
2016;38(4):388-392
- CountryChina
- Language:English
-
Abstract:
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.
