体外诱导毛囊Bulge细胞向皮脂腺细胞定向分化的研究
发布时间：2018-10-10 10:14
【摘要】： 毛囊是皮肤重要的附属器官,具有周期性生长的特点。目前已知毛囊干细胞位于外根鞘的隆突(bulge)区,研究证明毛囊bulge细胞具有多向分化的能力,不仅能分化形成毛囊,还能够形成皮脂腺和表皮角朊细胞。由于表皮、毛囊和皮脂腺关系密切,一般将三者统归为表皮-毛囊-皮脂腺单位(epidermo- piosebaceous unit)或表皮和毛囊皮脂腺单位(piosebaceous unit)。 皮脂腺(sebaceous gland)是皮肤重要的附属结构,具有抗炎、抵抗外来微生物的作用;它能够抗氧化,特别是抗紫外线的侵袭,对保持皮肤的完整和功能正常有着重要的作用。皮脂腺细胞的发育分化过程十分复杂,包括细胞形态的改变和基因表达的变化。过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor, PPARγ)在成脂细胞分化过程中起着关键的作用,它能有效促进脂质合成。在PPARγ的三种亚型中,PPARγ2与脂肪和皮脂腺细胞的分化尤其相关。PPARγ2是脂肪细胞分化过程中重要的调节因子,它可促使成纤维细胞或骨髓间充质干细胞向脂肪细胞分化。尽管多项证据表明皮脂腺的更新和维持来源于毛囊的Bulge细胞,但是,至今仍未在体外实验中观察到bulge细胞分化为皮脂腺细胞的直接证据。毛囊干细胞体外分离培养技术的相对滞后,影响了研究的进一步深入。我们在体外分离培养了毛囊bulge细胞,并在一定的条件下诱导bulge细胞向皮脂腺细胞定向分化,同时构建PPARγ2质粒,通过脂质体将其转染到bulge细胞中,观察它对bulge细胞定向分化为皮脂腺过程中的作用。主要结果如下: 1、毛囊bulge细胞的分离培养及生物学特性的研究 采用免疫组化及原位杂交技术,检测大鼠毛囊Bulge区细胞干细胞表面标志物和分化标志物的表达情况,运用显微分离及酶消化法分离并培养大鼠毛囊Bulge细胞,观察培养的毛囊bulge细胞体外生长特性,同时比较了不同培养体系对bulge细胞生物学特性的影响。结果发现:(1)干细胞表面标志物K19的表达主要位于毛囊的Bulge区,而该区不表达毛囊上皮的细胞分化标志K10。(2)经酶消化后的毛囊Bulge区组织易贴壁,24h后即有细胞爬出,沿Bulge区向外扩展生长,3-4天后细胞出现融合,
[Abstract]:Hair follicle is an important subsidiary organ of the skin, which has the characteristic of periodic growth. Hair follicle stem cells are known to be located in the protuberant (bulge) region of the outer root sheath. It has been demonstrated that hair follicle bulge cells have the ability to differentiate into hair follicles and sebaceous glands and epidermal keratinocytes. Because the epidermis, hair follicle and sebaceous gland are closely related, they are generally classified as epidermal hair follicle-sebaceous unit (epidermo- piosebaceous unit) or epidermis and hair follicle sebaceous gland unit (piosebaceous unit). Sebaceous gland (sebaceous gland) is an important subsidiary structure of skin, which has the function of anti-inflammation and resisting foreign microorganism, and it can resist the invasion of ultraviolet rays, and it has important function to maintain the integrity and function of skin. The process of development and differentiation of sebaceous gland cells is very complicated, including the changes of cell morphology and gene expression. Peroxisome proliferator activated receptor 纬 (peroxisome proliferator-activated receptor, PPAR 纬) plays a key role in the process of adipoblast differentiation, which can effectively promote lipid synthesis. Among the three subtypes of PPAR 纬, PPAR纬 2 is especially related to the differentiation of adipose and sebaceous gland cells. PPAR- 纬 2 is an important regulatory factor in adipocyte differentiation, which can induce fibroblasts or bone marrow mesenchymal stem cells to differentiate into adipocytes. Although many evidences suggest that the regeneration and maintenance of sebaceous glands originate from Bulge cells in hair follicles, there is no direct evidence that bulge cells differentiate into sebaceous gland cells in vitro. The technology of hair follicle stem cell isolation and culture in vitro is lagging behind, which affects the further research. Hair follicle bulge cells were isolated and cultured in vitro, and bulge cells were induced to differentiate into sebaceous gland cells under certain conditions. At the same time, PPAR 纬 2 plasmid was constructed and transfected into bulge cells by liposome. To observe its effect on the differentiation of bulge cells into sebaceous glands. The main results are as follows: 1. Isolation, culture and biological characteristics of hair follicle bulge cells Immunohistochemistry and in situ hybridization, The expression of stem cell surface markers and differentiation markers in Bulge region of rat hair follicle was detected. The Bulge cells of hair follicle were isolated and cultured by microdissection and enzyme digestion. The growth characteristics of cultured bulge cells were observed in vitro. The effects of different culture systems on the biological characteristics of bulge cells were also compared. The results showed that: (1) the expression of stem cell surface marker K19 was mainly located in the Bulge region of the hair follicle, but the cell differentiation marker of the hair follicle epithelium was not expressed in this area. (2) the tissue of the Bulge region of the hair follicle easily adhered to the wall 24 hours after the enzyme digestion. After 3 to 4 days of growth along the Bulge region, the cells were fused.
【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329
本文编号：2261396
[Abstract]:Hair follicle is an important subsidiary organ of the skin, which has the characteristic of periodic growth. Hair follicle stem cells are known to be located in the protuberant (bulge) region of the outer root sheath. It has been demonstrated that hair follicle bulge cells have the ability to differentiate into hair follicles and sebaceous glands and epidermal keratinocytes. Because the epidermis, hair follicle and sebaceous gland are closely related, they are generally classified as epidermal hair follicle-sebaceous unit (epidermo- piosebaceous unit) or epidermis and hair follicle sebaceous gland unit (piosebaceous unit). Sebaceous gland (sebaceous gland) is an important subsidiary structure of skin, which has the function of anti-inflammation and resisting foreign microorganism, and it can resist the invasion of ultraviolet rays, and it has important function to maintain the integrity and function of skin. The process of development and differentiation of sebaceous gland cells is very complicated, including the changes of cell morphology and gene expression. Peroxisome proliferator activated receptor 纬 (peroxisome proliferator-activated receptor, PPAR 纬) plays a key role in the process of adipoblast differentiation, which can effectively promote lipid synthesis. Among the three subtypes of PPAR 纬, PPAR纬 2 is especially related to the differentiation of adipose and sebaceous gland cells. PPAR- 纬 2 is an important regulatory factor in adipocyte differentiation, which can induce fibroblasts or bone marrow mesenchymal stem cells to differentiate into adipocytes. Although many evidences suggest that the regeneration and maintenance of sebaceous glands originate from Bulge cells in hair follicles, there is no direct evidence that bulge cells differentiate into sebaceous gland cells in vitro. The technology of hair follicle stem cell isolation and culture in vitro is lagging behind, which affects the further research. Hair follicle bulge cells were isolated and cultured in vitro, and bulge cells were induced to differentiate into sebaceous gland cells under certain conditions. At the same time, PPAR 纬 2 plasmid was constructed and transfected into bulge cells by liposome. To observe its effect on the differentiation of bulge cells into sebaceous glands. The main results are as follows: 1. Isolation, culture and biological characteristics of hair follicle bulge cells Immunohistochemistry and in situ hybridization, The expression of stem cell surface markers and differentiation markers in Bulge region of rat hair follicle was detected. The Bulge cells of hair follicle were isolated and cultured by microdissection and enzyme digestion. The growth characteristics of cultured bulge cells were observed in vitro. The effects of different culture systems on the biological characteristics of bulge cells were also compared. The results showed that: (1) the expression of stem cell surface marker K19 was mainly located in the Bulge region of the hair follicle, but the cell differentiation marker of the hair follicle epithelium was not expressed in this area. (2) the tissue of the Bulge region of the hair follicle easily adhered to the wall 24 hours after the enzyme digestion. After 3 to 4 days of growth along the Bulge region, the cells were fused.
【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2005
【分类号】：R329
【参考文献】
相关期刊论文 前6条
1 张勇,邹仲敏,郭朝华,周进明,王劲,范文辉,罗成基,程天民;MyoD基因诱导骨髓间充质干细胞分化为成肌细胞的实验研究[J];第三军医大学学报;2002年12期
2 张艺,杨恬;毛囊Bulge细胞培养与生物学特性的研究[J];第三军医大学学报;2004年16期
3 何诚,贺平,朱运松;nm23-H1-GFP融合蛋白在人肺癌细胞株中的表达及其对肿瘤细胞体外侵袭能力的影响[J];中国生物化学与分子生物学报;2000年01期
4 周红,郑江,鲁永玲,秦孝建,肖光夏;抗菌肽葛佬素在非洲绿猴肾细胞株COS-7中的表达及其活性的初步分析[J];中华烧伤杂志;2003年02期
5 赵宇,陆应麟,乔群,杜芝燕,徐元基,戚可名;绿色荧光蛋白基因逆转录病毒载体转染人骨髓间充质干细胞及表达的研究[J];中华实验外科杂志;2004年01期
6 游洪波,陈安民;增强型绿色荧光蛋白基因转染及TGFβ_3诱导前软骨干细胞向成软骨方向分化[J];中华小儿外科杂志;2004年06期
，本文编号：2261396
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2261396.html
最近更新
教材专著
