摘要
目的探讨改良的单酶消化法与组织贴块法相结合的培养雪旺细胞（SCs）的方法,并与经典植块法和普通消化复合植块法进行比较。方法取新生3～5 d的SD大鼠双侧坐骨神经和臂丛神经,分别采用经典植块法、普通消化复合植块法、改良消化复合植块法培养SCs。培养7 d后进行细胞消化和传代,在显微镜下计数并计算细胞总量。用S-100免疫荧光染色法检测所获细胞的纯度。结果改良消化复合植块法培养的SCs生长增殖速度最快,可获取（1.85±0.13）×10^6个细胞;普通消化复合植块法次之,可获取（1.10±0.10）×10^6个细胞;经典植块法所获SCs的生长增殖速度最慢,可获取（0.77±0.03）×10^6个细胞,3种方法比较差异均有统计学意义（P〈0.01）。S-100免疫荧光染色结果显示,改良消化复合植块法细胞纯度为（95.73±1.51）%,普通消化复合植块法为（84.66±2.68）%,经典植块法为（74.50±4.23）%,3种方法比较差异均有统计学意义（P〈0.01）。结论改良消化复合植块法可以在短时间内获得大量高纯度的SCs,为研究周围神经修复再生奠定良好的基础。
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） ×106; 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 explantsculture 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.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2016年第4期388-392,共5页
Acta Academiae Medicinae Sinicae
基金
协和青年科研基金(33320140118)~~
关键词
雪旺细胞
细胞培养
植块法
消化法
Schwann cells
cell culture
explants-culture method
enzyme digestion method
