CAJ | 학술논문

目的探索来源于成年大鼠周围神经体外培养的激活态雪旺细胞是否具有恶性潜能.方法从成年大鼠坐骨神经中分离、体外培养激活态雪旺细胞.通过对细胞形态连续动态观察、免疫学标志物鉴定、增殖率和纯度检测、迁移和侵袭能力检测、染色体核型分析及全基因组表达谱基因芯片、细胞因子抗体芯片检测,对激活态雪旺细胞是否具有恶性潜能进行评估.同时做自体体内接种试验,观察经体外激活、扩增后的激活态雪旺细胞接种到自体内是否可以致瘤.结果激活态雪旺细胞在体外培养形态较均一,无异质性变化.连续传代不丢失S100、P75LNGFR、GFAP等免疫标识物,激活前后迁移和侵袭能力略有增加,但远低于恶性细胞.染色体仍为2倍体.基因和蛋白表达存在动态平衡,第2、3代总体表达最高.动物自体接种可见细胞在体内非正常内环境下至少存活2个月,未见肿瘤生长.结论从成年大鼠周围神经分离、体外培养获得的激活态雪旺细胞,基本不具备恶性潜能,可用于填充人工神经导管修复周围神经缺损.
목적 탐색래원우성년대서주위신경체외배양적격활태설왕세포시부구유악성잠능.방법 종성년대서좌골신경중분리、체외배양격활태설왕세포.통과대세포형태련속동태관찰、면역학표지물감정、증식솔화순도검측、천이화침습능력검측、염색체핵형분석급전기인조표체보기인심편、세포인자항체심편검측,대격활태설왕세포시부구유악성잠능진행평고.동시주자체체내접충시험,관찰경체외격활、확증후적격활태설왕세포접충도자체내시부가이치류.결과 격활태설왕세포재체외배양형태교균일,무이질성변화.련속전대불주실S100、P75LNGFR、GFAP등면역표식물,격활전후천이화침습능력략유증가,단원저우악성세포.염색체잉위2배체.기인화단백표체존재동태평형,제2、3대총체표체최고.동물자체접충가견세포재체내비정상내배경하지소존활2개월,미견종류생장.결론 종성년대서주위신경분리、체외배양획득적격활태설왕세포,기본불구비악성잠능,가용우전충인공신경도관수복주위신경결손.
Objective To explore whether activated Schwann cells cultured and expanded from adult rat peripheral nerves have oncogenesis potentials. Methods After isolation from adult rat sciatic nerves, Schwann cells were activated and cultured in vitro. Through successive cell morphology observation, immunological marker identification, proliferation index and purity detection, migration and invasion assays, karyotype analysis, gene expression profiling and cytokine antibody chip analysis, oncogenesis potential of the activated Schwann cells was evaluated. In addition, sufficient activated Sehwann cells were autotransplanted into the axillary fossa to see whether they could produce tumors in vivo. Results Activated Schwann cells showed homogenous morphology which was identical to inactivated Schwann cells. Activation and successive passages did not influence the expression of Schwann cell markers such as S100, P75LNGFR and GFAP. After activation, the abilities of migration and invasion of Schwann cells were a little enhanced, but were much lower than that of malignant cells. The expressions of genes and proteins were dynamically balanced, while cells of the second and third passages had the highest expressions. Karyotype analysis of activated Schwann cells was normal. Autotransplantation test showed that activated Schwann cells survived for at least 2 months, and did not lead to tumor formation. Conclusion Activated Schwann cells, isolated from rat peripheral nerves and expanded in vitro, have no oncogenesis potential. These cells can be used to supplement artificial nerve conduits for nerve repair.[Key words ]Schwann cells; Cell culture techniques; Cell transplantation; Activated;Malignancy