Immortalized mesenchymal stem cells: an alternative to primary mesenchymal stem cells in neuronal differentiation and neuroregeneration associated studies.
Immortalized mesenchymal stem cells: an alternative to primary mesenchymal stem cells in neuronal differentiation and neuroregeneration associated studies.
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永生化间充质干细胞：神经元分化和神经再生相关研究中原代间充质干细胞的替代品
DOI：
10.1186/1423-0127-18-87
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发表时间：
2011-11-25
影响因子：
11
通讯作者：
Li T
中科院分区：
文献类型：
--
作者：
Gong M;Bi Y;Jiang W;Zhang Y;Chen L;Hou N;Liu Y;Wei X;Chen J;Li T
背景间充质干细胞（MSCs）可以在适当的细胞条件下诱导分化为神经细胞，并移植到脑损伤和神经退行性疾病动物模型中进行神经再生研究。与胚胎干细胞（ESCs）相比，间充质干细胞由于其有限的自我更新能力而容易老化和衰老。间充质干细胞的衰老严重影响了其作为细胞再生医学和组织工程工具的应用前景。本研究利用表达类人猿病毒40大T （SV40T）抗原的SSR#69逆转录病毒作为原代间充质干细胞的替代品，建立了可逆的永活间充质干细胞（IMSCs）系。方法利用表达猿猴病毒40大T （SV40T）抗原的逆转录病毒载体SSR#69构建IMSCs。采用流式细胞术检测细胞表面生成物。通过细胞生长曲线测定和中胚层三龄分化试验，研究IMSCs的增殖和分化潜能。体外检测IMSCs的神经元分化特征。在IMSCs移植前，我们首先排除其在裸鼠体内的致瘤性。在HIBD模型接受细胞移植治疗5周后进行Morris水迷宫试验和穿梭箱试验。结果本实验成功构建了可逆的间充质干细胞，其形态和细胞表面构成与原代间充质干细胞相似。与原代间充质干细胞相比，IMSCs具有更好的增殖和抗衰老能力，同时保持了多系分化能力。与来源于原代MSCs的神经样细胞相比，来源于IMSCs的神经样细胞具有相似的神经特异性基因表达、蛋白质表达模式和静息膜电位（RMP）。裸鼠皮下注射IMSCs后未见肿块形成。IMSCs对HIBD大鼠学习能力和空间记忆的改善作用与原代MSCs相同。结论simscs不仅保留了原代MSCs的特征，而且具有高增殖和抗衰老的能力。在体外完全可以诱导间充质干细胞分化为神经元细胞，在体内不发生肿瘤的情况下代替原代间充质干细胞进行细胞移植治疗。这种稳定的细胞系在神经元分化和神经再生相关研究中作为MSCs的替代品尤其有用和有价值。
BackgroundMesenchymal stem cells (MSCs) can be induced to differentiate into neuronal cells under appropriate cellular conditions and transplanted in brain injury and neurodegenerative diseases animal models for neuroregeneration studies. In contrast to the embryonic stem cells (ESCs), MSCs are easily subject to aging and senescence because of their finite ability of self-renewal. MSCs senescence seriously affected theirs application prospects as a promising tool for cell-based regenerative medicine and tissue engineering. In the present study, we established a reversible immortalized mesenchymal stem cells (IMSCs) line by using SSR#69 retrovirus expressing simian virus 40 large T (SV40T) antigen as an alternative to primary MSCs.MethodsThe retroviral vector SSR#69 expressing simian virus 40 large T (SV40T) antigen was used to construct IMSCs. IMSCs were identified by flow cytometry to detect cell surface makers. To investigate proliferation and differentiation potential of IMSCs, cell growth curve determination and mesodermal trilineage differentiation tests were performed. Neuronal differentiation characteristics of IMSCs were detectedin vitro. Before IMSCs transplantation, we excluded its tumorigenicity in nude mice firstly. The Morris water maze tests and shuttle box tests were performed five weeks after HIBD models received cells transplantation therapy.ResultsIn this study, reversible IMSCs were constructed successfully and had the similar morphology and cell surface makers as primary MSCs. IMSCs possessed better ability of proliferation and anti-senescence compared with primary MSCs, while maintained multilineage differentiation capacity. Neural-like cells derived from IMSCs had similar expressions of neural-specific genes, protein expression patterns and resting membrane potential (RMP) compared with their counterparts derived from primary MSCs. There was no bump formation in nude mice subcutaneously injected with IMSCs. IMSCs played same role as primary MSCs to improve learning ability and spatial memory of HIBD rats.ConclusionsIMSCs not only retain their features of primary MSCs but also possess the ability of high proliferation and anti-senescence. IMSCs can definitely be induced to differentiate into neuronal cellsin vitroand take the place of primary MSCs for cell transplantation therapy without tumorigenesisin vivo. The stable cell line is particularly useful and valuable as an alternative to MSCs in neuronal differentiation and neuroregeneration associated studies.