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诱导大鼠骨髓间充质干细胞分化为唾液腺腺泡样细胞的实验研究(1)
http://www.100md.com 2012年3月1日 梁亮,孙沫逸,李建虎,杨永勤,申志远,苏忠平
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     [摘要]目的:探讨在共培养系统下大鼠骨髓间充质干细胞分化为唾液腺腺泡样细胞的实验。方法:以纯化1代SD大鼠颌下腺腺泡细胞和3代骨髓间充质干细胞作为共培养实验对象。实验分组:含唾液腺培养液的共培养组;含10%FBS、 DMEM/F12的共培养组;含唾液腺培养液的非共培养组;含10%FBS、DMEM/F12的非共培养组。培养1个月经α-淀粉酶(α-amylase)免疫组化染色各组的MSCs,计算阳性细胞数得出MSCs的转化率,并且通过光镜和电镜鉴定细胞形态变化。结果:各组诱导的MSCs经α-amylase染色,共培养组阳性细胞数较非共培养组多(P<0.05),且诱导成功的细胞在镜下形态类似于唾液腺腺泡细胞。结论:在共培养条件下成功实现了MSCs向 SGCs的形态学转化,唾液腺专用培养液能够提高SGCs的诱导效率。

    [关键词]骨髓间充质干细胞;唾液腺腺泡细胞;共培养;诱导

    [中图分类号]Q813.1 [文献标识码]A [文章编号]1008-6455(2012)03-0419-03

    The research of rat bone marrow mesenchymal stem cells differentiation into salivary gland acinar-like cells

    LIANG Liang,SUN Mo-yi,LI Jian-hu,YANG Yong-qin,SHEN Zhi-yuan,SU Zhong-ping

    (Department of Oral and Maxillofacial Surgery,School of Stomatology,The Fourth Militiary Medical University,Xi'an 710032,Shaanxi,China)

    Abstact: Objective To explore the possibility of rat bone marrow mesenchymal stem cells(MSCs) differentiation into salivary gland acinar-like cells induced by co-culture with salivary gland acinar cells(SGCS) in vitro. Methods SGCs of passage 1 and MSCs of passage 3 were taken as subject investigated.Following groups were allocated:co-culture with salivary gland medium group;co-culture with 10% FBS DMEM/F12 group;salivary gland medium group;group with 10% FBS DMEM/F12.MSCs of every group were identified by immunohistochemiscal analysis withα-amylase After a month cultivation.MSCs conversion rate is attained by calculation of the number of positive cells.By the optical microscope and scanning electron microscope observes the morphological changes of cells. Results Compared with none-co-cultured groups,Most cells in co-culture groups were stained by α-amylase(P<0.05). Conclusions The cells of success induction morphology which similar to salivary gland acinar cells at the optical microscope and scanning electron microscope display.MSCs could differentiate into SGs at phenotype and molecule level by co-culture of SGs in vitro.Changing the experimental conditions and can improve induction efficiency.

    Key words:bone marrow mesenchymal stem cell;salivary gland acinar cell;co-culture;induction ......

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