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In vitro differentiation of human adipose-derived adult stromal cells into neuron-like cells in hippocampal astrocyte conditioned medium

机译:人脂肪来源的成人基质细胞在海马星形胶质细胞条件培养基中的体外分化为神经元样细胞

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摘要

BACKGROUND: At present, researches on differentiating from human adipose-derived adult stromal cells (hADASC) to neuron-like cells are focus on inducing by artificial-synthetic compound solution;however,hippocampal astrocyte conditioned medium(HCAM)can induce in vitro differentiation from hADASC to neuron-like cells is still unclear.OBJECTIVE:To observe whether HCAM can induce in vitro differentiation from hADASC to neuron-like cells.DESIGN:Randomized control study.SETTING:Department of Neurology,Taixing People's Hospital;Central Laboratory,North China Coal Medical College.MATERIALS:Donor of adipose tissue was donated by female volunteers suffering from caesarean section in the department of obstetrics & gynecology in our hospital and aged 20-35 years. Adipose tissue was collected from subcutaneous tissue of abdomen during the operation.In addition.8 male newborn Wistar rats within 24 hours with average body mass of 20 g were provided by Animal Institute of Chinese Academy of Medical Sciences.Rabbit-anti-human Nestin polyclonal antibody.Rabbit-anti-human glial fibriliary acidic protein (GFAP)polyclonal antibody, rabbit-anti-human neuro-specific enolase polyclonal antibody and mouse-anti-human microtubal associated protein 2(MAP-2)polyclonal antibody were provided by Wuhan Boster Company.METHODS:The experiment was carried out in the Central Laboratory of North China Coal Medical College from October 2004 to June 2005.hADASC was cultured with HCAM and its growth and morphological changes were observed under inverted phase contrast microscope.Immunocytochemistry.immunofluorescence and Western blotting were used to evaluate the expressions of Nestin,which was a specific sign of nerve precursor,neuro-specific enolase and MAP-2,which was a specific sign of nerve cell,and GFAP,which was a specific sign of neuroglial cells.MAIN OUTCOME MEASURES:Nestin,which was a specific sign of nerve precursor,neuro-specific enolase and MAP-2,which was a specific sign of nerve cell,and GFAP,which was a specific sign of neuroglial cells.RESULTS:On the 3rd day of culture.partial hADASC started deformation from slender shuttle-shape cells to neuron-like cells. It suggested that cells stretched out apophysis. Which were mainly double-pole or multiple-pole cells.Five days later,immunohistochemical detection suggested that expression of Nestin(10.5±0.037)was found out in cells;meanwhile,expressions of GFAP(38.4±0.052)and neuro-specific enolase(NSE)(15.7±0.023)were also found out in cells;however,expression of MAP-2 was not observed.Western blot indicated that,5 days after effect of HCAM,Nestin was found out in hADASC;meanwhile,expressions of GFAP and neuro-specific enolase were also found out:however,expression of MAP-2 was not observed.CONCLUSION:HCAM can induce the differentiation from hADASC to neuron-like cells in vitro.
机译:背景:目前,关于从人脂肪来源的成体基质细胞(hADASC)向神经元样细胞分化的研究,主要是通过人工合成的复合溶液诱导;然而,海马星形胶质细胞条件培养基(HCAM)可以诱导人成体细胞的体外分化。目的:观察HCAM是否可以诱导hADASC向神经元样细胞的体外分化。目的:随机对照研究。单位:泰兴市人民医院神经内科;华北中心材料:脂肪组织的捐赠是由我院妇产科剖腹产,年龄在20-35岁之间的女性志愿者捐赠的。手术过程中从腹部皮下组织中收集脂肪组织。此外,中国医学科学院动物研究所提供了24只24小时内平均体重为20 g的雄性新生Wistar大鼠。兔抗人巢蛋白多克隆抗体武汉博斯特提供了兔抗人神经胶质纤维酸性蛋白(GFAP)多克隆抗体,兔抗人神经特异性烯醇化酶多克隆抗体和小鼠抗人微管相关蛋白2(MAP-2)多克隆抗体公司方法:本实验于2004年10月至2005年6月在华北煤炭医学院中心实验室进行。用HCAM培养hADASC,并在反相显微镜下观察其生长和形态变化。免疫细胞化学,免疫荧光和Western用印迹法检测Nestin的表达,Nestin是神经前体的特异信号,神经特异性烯醇化酶和MAP-2是一种神经元。主要观察指标:Nestin,神经前体的特异性体征,神经特异性烯醇酶和MAP-2,是神经细胞的特异性体征。结果:在培养的第3天,部分hADASC开始从细长的穿梭形细胞变形为神​​经元样细胞。这表明细胞伸展了骨质疏松症。五天后,免疫组织化学检测发现在细胞中发现了巢蛋白(10.5±0.037)的表达;同时,表达了GFAP(38.4±0.052)和神经特异性烯醇化酶(在细胞中也发现了NSE(15.7±0.023);但未观察到MAP-2的表达。Westernblot显示,在HCAM作用5天后,hADASC中发现了Nestin;同时GFAP和GFAP的表达也升高。还发现了神经特异性烯醇化酶:但是未观察到MAP-2的表达。结论:HCAM可以诱导hADASC向神经元样细胞的分化。

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  • 来源
    《中国神经再生研究(英文版)》 |2006年第7期|581-584|共4页
  • 作者单位

    Department of Neurology,Taixing People's Hospital,Taixing 225400,Jiangsu Province,China;

    Department of Neurology,Taixing People's Hospital,Taixing 225400,Jiangsu Province,China;

    Department of Neurology,Taixing People's Hospital,Taixing 225400,Jiangsu Province,China;

    Department of Neurology,Taixing People's Hospital,Taixing 225400,Jiangsu Province,China;

    Department of Neurology,Taixing People's Hospital,Taixing 225400,Jiangsu Province,China;

    Department of Neurology,Affiliated Hospital of North China Coal Medical College,Tangshan 063000,Hebei Province,China;

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  • 正文语种 chi
  • 中图分类 神经病学与精神病学;
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  • 入库时间 2022-08-19 03:44:52
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