CAJ | 학술논문

目的探讨小鼠非黏附骨髓间充质干细胞(non-adherent bone marrow-derived mesen-chymal stem cell,NA-BM-MSC)在体外、体内尤其是缺血损伤的脑内向神经元样细胞分化的能力.方法传代纯化后的NA-BM-MSC经表皮生长因子和碱性成纤维细胞生长因子诱导后,通过甲苯胺蓝染色和免疫细胞化学染色检测神经元特异性核蛋白和神经中间丝蛋白的表达情况;将来自β-半乳糖苷酶转基因小鼠的NA-BM-MSC在立体定位仪下移植到小鼠缺血损伤的脑内,8周后通过LacZ组织化学染色和免疫组织化学染色检测供体细胞在缺血脑内微环境中存活以及神经元特异性核蛋白的表达情况. 结果 NA-BM-MSC经过诱导能够表达神经元特异性核蛋白和神经中间丝蛋白并有尼氏体形成;移植到损伤脑内后,LacZ组织化学染色发现在缺血侧供体细胞能够存活,免疫组织化学单、双染显示在缺血坏死区及周围都检测到了β-半乳糖苷酶阳性的供体细胞,其中的部分阳性细胞还同时表达神经元特异性核蛋白. 结论 NA-BM-MSC在体外能够分化成神经元样细胞,在小鼠缺血损伤的脑组织内也能够存活、迁移,部分细胞还能分化为神经元样细胞.
목적 탐토소서비점부골수간충질간세포(non-adherent bone marrow-derived mesen-chymal stem cell,NA-BM-MSC)재체외、체내우기시결혈손상적뇌내향신경원양세포분화적능력.방법 전대순화후적NA-BM-MSC경표피생장인자화감성성섬유세포생장인자유도후,통과갑분알람염색화면역세포화학염색검측신경원특이성핵단백화신경중간사단백적표체정황;장래자β-반유당감매전기인소서적NA-BM-MSC재입체정위의하이식도소서결혈손상적뇌내,8주후통과LacZ조직화학염색화면역조직화학염색검측공체세포재결혈뇌내미배경중존활이급신경원특이성핵단백적표체정황. 결과 NA-BM-MSC경과유도능구표체신경원특이성핵단백화신경중간사단백병유니씨체형성;이식도손상뇌내후,LacZ조직화학염색발현재결혈측공체세포능구존활,면역조직화학단、쌍염현시재결혈배사구급주위도검측도료β-반유당감매양성적공체세포,기중적부분양성세포환동시표체신경원특이성핵단백. 결론 NA-BM-MSC재체외능구분화성신경원양세포,재소서결혈손상적뇌조직내야능구존활、천이,부분세포환능분화위신경원양세포.
Objecttve To determine whether non-adherent bone marrow-derived mesenchymal stem cells (NA-BM-MSC) can differentiate into neuron-like cells in vitro and in vivo,especially in ischemic brain of mice. Methods NA-BM-MSC of mouse were indueed with conditioned medium containing epidermal growth factor and basic-fibroblast growth factor and demonstrated by immunocytochemistry for the expression of neuron specific nuclear protein and neurofilament-200. NA-BM-MSC from β-galactosidase transgenic mice constitutively expressing β-galactosidase were transplanted into the mice model of middle cerebral artery occlusion.LacZ staining and immunobistochemistry staining were performed to detect the survival,distribution and the differentiation of the donor cells in the ischemic brain. Results NA-BM-MSC can be induced into neuron specific nuclear protein and neurofilament-200 positive cells in vitro.LacZ staining showed that NA-BM-MSC can survive in ischemic brain and express β-galactosidase.Also,numbers of the neuron specific nuclear protein positive cells in β-galactosidase-positive cells were detected in the ischemic brain with double immunohistochemistry staining. Conclusions NA-BM-BMC can be induced to differentiate into neuron-like cells in vitro,and also can differentiate into neuron-like cells in ischemic brain tissue of the mice.