中华围产医学杂志
中華圍產醫學雜誌
중화위산의학잡지
CHINESE JOURNAL OF PERINATAL MEDICINE
2010年
2期
123-127
,共5页
王燕%陈燕玲%贲晓明%周晓玉%苗登顺
王燕%陳燕玲%賁曉明%週曉玉%苗登順
왕연%진연령%분효명%주효옥%묘등순
间质干细胞%细胞分化%神经元%脑缺血%间质干细胞移植
間質榦細胞%細胞分化%神經元%腦缺血%間質榦細胞移植
간질간세포%세포분화%신경원%뇌결혈%간질간세포이식
Mesenchymal stem cells%Cell differentiation%Neurons%Brain ischemia%Mesenchymal stem cell transplantation
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.
目的 探討小鼠非黏附骨髓間充質榦細胞(non-adherent bone marrow-derived mesen-chymal stem cell,NA-BM-MSC)在體外、體內尤其是缺血損傷的腦內嚮神經元樣細胞分化的能力.方法 傳代純化後的NA-BM-MSC經錶皮生長因子和堿性成纖維細胞生長因子誘導後,通過甲苯胺藍染色和免疫細胞化學染色檢測神經元特異性覈蛋白和神經中間絲蛋白的錶達情況;將來自β-半乳糖苷酶轉基因小鼠的NA-BM-MSC在立體定位儀下移植到小鼠缺血損傷的腦內,8週後通過LacZ組織化學染色和免疫組織化學染色檢測供體細胞在缺血腦內微環境中存活以及神經元特異性覈蛋白的錶達情況. 結果 NA-BM-MSC經過誘導能夠錶達神經元特異性覈蛋白和神經中間絲蛋白併有尼氏體形成;移植到損傷腦內後,LacZ組織化學染色髮現在缺血側供體細胞能夠存活,免疫組織化學單、雙染顯示在缺血壞死區及週圍都檢測到瞭β-半乳糖苷酶暘性的供體細胞,其中的部分暘性細胞還同時錶達神經元特異性覈蛋白. 結論 NA-BM-MSC在體外能夠分化成神經元樣細胞,在小鼠缺血損傷的腦組織內也能夠存活、遷移,部分細胞還能分化為神經元樣細胞.
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.