Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica

Acta Academiae Medicinae Sinica ›› 2016, Vol. 38 ›› Issue (4): 388-392.doi: 10.3881/j.issn.1000-503X.2016.04.004

• Orginal Article • Previous Articles     Next Articles

Culturing Schwann Cells from Neonatal Rats by Improved Enzyme Digestion Combined with Explants-culture Method

Di LIU1, Xiao-chun LIANG1(), Hong ZHANG2   

  1. 1Department of Traditional Chinese Medicine,PUMC Hospital,CAMS and PUMC,Beijing 100730,China
    2Department of Cell Resource Center,Institute of Basic Medical Science,CAMS and PUMC,Beijing 100005,China
  • Received:2015-07-21 Online:2016-08-20 Published:2016-08-20
  • Supported by:
    Supported by the Youth Scientific Research Foundation of PUMC (33320140118)

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)×106;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)×106;the SCs of the explants-culture method group grew slowest and the total number of cells obtained was (0.77±0.03)×106.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.

Key words: Schwann cells, cell culture, explants-culture method, enzyme digestion method

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