猪小肠黏膜下层与嗅鞘细胞生物相容性实验

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摘要目的：探讨以小肠黏膜下层（SIS）作为生物支架对嗅鞘细胞（OECs）增殖的生物相容性。方法：用Abraham法制取猪小肠黏膜下层，将小肠黏膜下层与纯化的嗅鞘细胞复合培养作为实验组，单纯嗅鞘细胞培养作为对照组；定期进行倒置显微镜、HE染色、扫描电镜观察细胞与材料的黏附生长情况，及CCK8增殖实验定量检测细胞增殖情况。结果：实验组小肠黏膜下层与嗅鞘细胞复合培养后，倒置显微镜下可见嗅鞘细胞在24 h内即可贴壁生长；3～5 d后可见小肠黏膜下层边缘嗅鞘细胞密集，黏附良好，形态以梭形细胞、三极细胞为主；不同时间点HE染色、电镜下可见嗅鞘细胞与猪小肠黏膜下层附着紧密，细胞生长良好。CCK8细胞增殖定量检测发现，实验组细胞7～9 d后达平稳水平，11 d后略下降；前5 d实验组细胞增殖能力低于对照组，差异有统计学意义（均P＜0.05），培养至第7天2组细胞增殖能力差异无统计学意义（均P＞0.05）。结论：未发现小肠黏膜下层对嗅鞘细胞增殖有抑制作用，两者间生物相容性良好，有望用作生物支架供嗅鞘细胞黏附生长。

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	李玉安1
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	徐华梓1
	林大木1
	吴志鹏1

关键词 ：嗅鞘细胞, 小肠黏膜下层, 支架, 脊髓损伤

Abstract：Objective: To explore the biocompatibility of small intestinal submucosa as a biological scaffold on the proliferation of olfactory ensheathing cells. Methods: The porcine small intestinal submucosa (SIS) was made according to Abraham’s methods. seeding the OECs on the porcine SIS for experimental group (SIS+OECs), OECs cultivating alone as control group. Then OECs’ growth, adhesion and proliferation of OECs on SIS was examined by HE stained and scanning electron microscope at different time during the cultivation, and the proliferation of OECs was examined with CCK8 as well. Results: OECs could attach to SIS within 24 hours after seeded on the SIS. After being cultured 3 to 5 days, OECs were intensive on the edge of SIS, well adhered to it and the cells were most fusiform and tripolar in shape. The combination of SIS and OECs was very excellently examinated by scanning electron microscope and Haematoxylin and eosin stained. And CCK8 measurement also revealed that, the quantity of OECs reached a stable level at 7 to 9 day and this level would not decreased until 11 d. Compared with these in contrast group, the proliferation of OECs on SIS had obvious difference during the former 5 days (P<0.05) but there was no significant difference after 7 day (P>0.05). Conclusion: Porcine SIS has good biocompatibility with rat OECs, although there is no evidence showing SIS promoting OECs’ proliferation. SIS can expectedly be used in repair of spinal cord injury as a scaffold where OECs may grow and adhere.

Key words： olfactory ensheathing cells small submucosa intestinal scaffold spinal cord injury

收稿日期: 2014-07-31

基金资助:温州市科技局对外合作项目（H20080031）。

通讯作者: 徐华梓，教授，主任医师，博士生导师，Email：spinexu@163.com。

作者简介: 李玉安（1983-），男，浙江温州人，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2015/V45/I9/656

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