白介素6类细胞因子对成人成肌细胞增殖和分化影响的实验研究
发布时间：2018-07-15 14:06
【摘要】：白介素6类细胞因子家族,调控着细胞的分化、存活、凋亡和增殖,参与多种复杂的细胞过程如抗炎、造血和神经分化等,是一类重要的细胞因子。白介素6(Interleukin-6,IL-6)、白血病抑制因子(Leukemia inhibitory factor,LIF)和睫状神经营养因子(Ciliary neurotrophic factor, CNTF)是其中较重要的三种因子,它们对于血液系统和神经系统的影响已经被广泛研究,但在人源成体成肌细胞的增殖和分化中的作用还未见报导。而成肌细胞做为一种能够提供丰富细胞来源的细胞群体,除了在肌肉萎缩等疾病中有重要作用外,由于它们可以自体取材,因此在以神经细胞变性为特征的中枢神经系统退行性疾病如帕金森病中,也具有较大的应用价值。 我们研究所前期的工作已经证实,一定浓度的CNTF可以抑制成肌细胞向成熟骨骼肌分化,而形成具有向多种细胞类型包括神经元分化潜能的更加原始的干细胞。因此,在本实验中,将应用IL-6、LIF和CNTF三种因子处理成人骨骼肌来源的单克隆化的成肌细胞,观察并比较三种因子对成人成肌细胞增值和分化的影响,并初步探讨它们的作用机制,以证实这些细胞因子是否是一类新的成肌细胞增殖和分化调控分子。同时进行成人成肌细胞脑移植的初步实验,验证成肌细胞做为中枢神经系统疾病移植治疗的替代细胞的可能性。 1、原代培养成人成肌细胞体外增殖、传代、冻存与复苏 连续贴壁法筛选获得的成人成肌细胞纯度不高,多混杂有成纤维细胞的干扰;应用96孔培养板进行的细胞单克隆化,使成肌细胞纯度明显增高,成肌细胞标志蛋白—结蛋白(Desmin)染色结果表明,由此获得的成肌细胞是Desmin阳性、纯度一致、稳定的细胞群体。 2、IL-6、LIF、CNTF细胞因子对成人成肌细胞增殖的影响 细胞增殖曲线和流式细胞术检测结果稍有不同,总结两种方法进行的细胞增殖研究,可明确10ng/ml LIF能够明显促进成人成肌细胞的增殖。而IL-6和CNTF的作用不确切。
[Abstract]:Interleukin-6 family of cytokines, which regulate cell differentiation, survival, apoptosis and proliferation, participate in many complex cellular processes, such as anti-inflammatory, hematopoietic and neural differentiation, are important cytokines. Interleukin 6 (IL 6), leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) are three of them. Their effects on blood system and nervous system have been widely studied. However, the role of myoblasts in the proliferation and differentiation of human adult myoblasts has not been reported. And myoblasts, as a group of cells that provide a rich source of cells, in addition to playing an important role in diseases such as muscular atrophy, because they can take materials from themselves, Therefore, neurodegenerative diseases such as Parkinson's disease, which are characterized by neuronal degeneration, also have great application value. Our previous work has confirmed that CNTF at a certain concentration can inhibit the differentiation of myoblasts into mature skeletal muscles and form more primitive stem cells with the potential to differentiate into a variety of cell types including neurons. Therefore, in this experiment, IL-6 LIF and CNTF were used to treat adult skeletal muscle derived monoclonal myoblasts, to observe and compare the effects of the three factors on the proliferation and differentiation of adult myoblasts, and to explore their mechanism. To verify whether these cytokines are a new class of myoblast proliferation and differentiation regulation molecules. At the same time, a preliminary experiment of adult myoblast brain transplantation was carried out. To verify the possibility of myoblasts as substitute cells for transplantation treatment of central nervous system diseases. 1. Primary cultured adult myoblasts proliferate and subculture in vitro. The purity of adult myoblasts selected by continuous adherent method of cryopreservation and resuscitation was not high, and most of them were mixed with fibroblasts. The purity of myoblast was obviously increased by using the McAb of 96-well culture plate. The result of desmin staining showed that the myoblast was Desmin positive and the purity was the same. The effect of IL-6 LIF-CNTF cytokines on the proliferation of adult myoblasts The results of cell proliferation curve and flow cytometry were slightly different. It is clear that 10ng/ml LIF can significantly promote the proliferation of adult myoblasts. But the effect of IL-6 and CNTF is not exact.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329
本文编号：2124327
[Abstract]:Interleukin-6 family of cytokines, which regulate cell differentiation, survival, apoptosis and proliferation, participate in many complex cellular processes, such as anti-inflammatory, hematopoietic and neural differentiation, are important cytokines. Interleukin 6 (IL 6), leukemia inhibitory factor (LIF) and ciliary neurotrophic factor (CNTF) are three of them. Their effects on blood system and nervous system have been widely studied. However, the role of myoblasts in the proliferation and differentiation of human adult myoblasts has not been reported. And myoblasts, as a group of cells that provide a rich source of cells, in addition to playing an important role in diseases such as muscular atrophy, because they can take materials from themselves, Therefore, neurodegenerative diseases such as Parkinson's disease, which are characterized by neuronal degeneration, also have great application value. Our previous work has confirmed that CNTF at a certain concentration can inhibit the differentiation of myoblasts into mature skeletal muscles and form more primitive stem cells with the potential to differentiate into a variety of cell types including neurons. Therefore, in this experiment, IL-6 LIF and CNTF were used to treat adult skeletal muscle derived monoclonal myoblasts, to observe and compare the effects of the three factors on the proliferation and differentiation of adult myoblasts, and to explore their mechanism. To verify whether these cytokines are a new class of myoblast proliferation and differentiation regulation molecules. At the same time, a preliminary experiment of adult myoblast brain transplantation was carried out. To verify the possibility of myoblasts as substitute cells for transplantation treatment of central nervous system diseases. 1. Primary cultured adult myoblasts proliferate and subculture in vitro. The purity of adult myoblasts selected by continuous adherent method of cryopreservation and resuscitation was not high, and most of them were mixed with fibroblasts. The purity of myoblast was obviously increased by using the McAb of 96-well culture plate. The result of desmin staining showed that the myoblast was Desmin positive and the purity was the same. The effect of IL-6 LIF-CNTF cytokines on the proliferation of adult myoblasts The results of cell proliferation curve and flow cytometry were slightly different. It is clear that 10ng/ml LIF can significantly promote the proliferation of adult myoblasts. But the effect of IL-6 and CNTF is not exact.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329
【共引文献】
相关博士学位论文 前1条
1 袁颖;牛膝多肽促神经生长作用及其相关机制的研究[D];苏州大学;2009年
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