【摘要】：白介素6類細(xì)胞因子家族,調(diào)控著細(xì)胞的分化、存活、凋亡和增殖,參與多種復(fù)雜的細(xì)胞過程如抗炎、造血和神經(jīng)分化等,是一類重要的細(xì)胞因子。白介素6(Interleukin-6,IL-6)、白血病抑制因子(Leukemia inhibitory factor,LIF)和睫狀神經(jīng)營養(yǎng)因子(Ciliary neurotrophic factor, CNTF)是其中較重要的三種因子,它們對于血液系統(tǒng)和神經(jīng)系統(tǒng)的影響已經(jīng)被廣泛研究,但在人源成體成肌細(xì)胞的增殖和分化中的作用還未見報(bào)導(dǎo)。而成肌細(xì)胞做為一種能夠提供豐富細(xì)胞來源的細(xì)胞群體,除了在肌肉萎縮等疾病中有重要作用外,由于它們可以自體取材,因此在以神經(jīng)細(xì)胞變性為特征的中樞神經(jīng)系統(tǒng)退行性疾病如帕金森病中,也具有較大的應(yīng)用價(jià)值。 我們研究所前期的工作已經(jīng)證實(shí),一定濃度的CNTF可以抑制成肌細(xì)胞向成熟骨骼肌分化,而形成具有向多種細(xì)胞類型包括神經(jīng)元分化潛能的更加原始的干細(xì)胞。因此,在本實(shí)驗(yàn)中,將應(yīng)用IL-6、LIF和CNTF三種因子處理成人骨骼肌來源的單克隆化的成肌細(xì)胞,觀察并比較三種因子對成人成肌細(xì)胞增值和分化的影響,并初步探討它們的作用機(jī)制,以證實(shí)這些細(xì)胞因子是否是一類新的成肌細(xì)胞增殖和分化調(diào)控分子。同時(shí)進(jìn)行成人成肌細(xì)胞腦移植的初步實(shí)驗(yàn),驗(yàn)證成肌細(xì)胞做為中樞神經(jīng)系統(tǒng)疾病移植治療的替代細(xì)胞的可能性。 1、原代培養(yǎng)成人成肌細(xì)胞體外增殖、傳代、凍存與復(fù)蘇 連續(xù)貼壁法篩選獲得的成人成肌細(xì)胞純度不高,多混雜有成纖維細(xì)胞的干擾;應(yīng)用96孔培養(yǎng)板進(jìn)行的細(xì)胞單克隆化,使成肌細(xì)胞純度明顯增高,成肌細(xì)胞標(biāo)志蛋白—結(jié)蛋白(Desmin)染色結(jié)果表明,由此獲得的成肌細(xì)胞是Desmin陽性、純度一致、穩(wěn)定的細(xì)胞群體。 2、IL-6、LIF、CNTF細(xì)胞因子對成人成肌細(xì)胞增殖的影響 細(xì)胞增殖曲線和流式細(xì)胞術(shù)檢測結(jié)果稍有不同,總結(jié)兩種方法進(jìn)行的細(xì)胞增殖研究,可明確10ng/ml LIF能夠明顯促進(jìn)成人成肌細(xì)胞的增殖。而IL-6和CNTF的作用不確切。
[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.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R329

【共引文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 袁穎;牛膝多肽促神經(jīng)生長作用及其相關(guān)機(jī)制的研究[D];蘇州大學(xué);2009年

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