残耳软骨细胞诱导脂肪来源干细胞体内软骨的形成

发布时间：2018-02-03 00:49

  本文关键词： 软骨细胞 脂肪类 干细胞 组织工程 脂肪干细胞 残耳软骨细胞 脂肪来源干细胞 软骨形成 共移植 诱导 体内　出处：《中国组织工程研究》2017年21期 　论文类型：期刊论文

【摘要】：背景:先天性小耳畸形残耳软骨细胞从细胞数量和质量上难以作为种子细胞构建出正常人耳郭大小的同质耳软骨支架。目的:探讨残耳软骨细胞能否在裸鼠体内非软骨环境下模拟软骨诱导微环境,促进脂肪来源的干细胞向软骨分化并形成组织工程软骨,从而为解决组织工程化人耳郭软骨支架制备做基础准备工作。方法:(1)实验分为4组:第2代残耳软骨细胞与第3代脂肪来源的干细胞以3∶7比例混合作为共移植组,单纯残耳软骨细胞作为阳性对照组(残耳软骨细胞组),单纯脂肪来源的干细胞作为阴性对照组(脂肪干细胞组),以上3组接种细胞终浓度为5.0×10~(10) L~(-1),低浓度残耳软骨细胞对照组细胞终浓度为1.5×10~(10) L~(-1);(2)按照实验分组将0.2 mL细胞-Pluronic-F127复合物注射到裸鼠背部皮下,体内培养8周后对新生组织进行大体观察、湿质量测量、糖胺多糖含量测定、组织学及免疫组化染色检测。结果与结论:(1)共移植组平均湿质量达到残耳软骨细胞组的80%以上;低浓度残耳软骨细胞对照组平均湿质量低于残耳软骨细胞组的30%;(2)共移植组和残耳软骨细胞组的平均湿质量和糖胺多糖平均含量均显著高脂肪干细胞组和低浓度残耳软骨细胞对照组(P0.05);(3)组织学染色:共移植组、残耳软骨细胞组与低浓度残耳软骨细胞对照组标本均有成熟的软骨陷窝形成,低浓度残耳软骨细胞对照组软骨陷窝松散排列不均,胞外基质着色淡;脂肪干细胞组为纤维样组织,未见软骨陷窝形成;(4)Ⅱ型胶原免疫组化染色:共移植组、残耳软骨细胞组与低浓度残耳软骨细胞对照组可见成熟软骨陷窝周围有不同程度的棕黄色沉淀即Ⅱ型胶原表达;脂肪干细胞组未见Ⅱ型胶原表达;(5)结果提示,残耳软骨细胞能够在裸鼠体内非软骨环境下模拟软骨诱导微环境,促进脂肪干细胞向软骨分化并生成组织工程软骨。
[Abstract]:Background: the quantity and quality of residual auricular chondrocytes in congenital microauricular malformation are difficult to be used as seed cells to construct homogenous auricular cartilage scaffolds of normal human auricle. To investigate whether residual ear chondrocytes can mimic cartilage induced microenvironment in nude mice in non-cartilage environment. Promote adipose derived stem cells to differentiate into cartilage and form tissue engineered cartilage. So as to solve the tissue engineering of human auricle cartilage scaffolds to prepare the basic work. Methods: 1). The experiment was divided into four groups: the second generation of residual ear chondrocytes and the third generation of adipose derived stem cells were mixed in the ratio of 3: 7 as co-transplantation group. Pure residual ear chondrocytes were used as positive control group (residual ear chondrocyte group) and adipose derived stem cells as negative control group (adipose stem cell group). The final concentration of inoculated cells was 5.0 脳 10 ~ (10) L ~ (-1) in the above three groups and 1.5 脳 10 ~ (10) L ~ (-1) in the control group with low concentration of residual ear chondrocytes. (2) 0.2 mL cell line -Pluronic-F127 complex was injected subcutaneously into the back of nude mice according to the experimental group. After 8 weeks of culture in vivo, the newborn tissues were observed and the wet mass was measured. Results and conclusion the average wet mass of the co-transplantation group was more than 80% of that of the residual ear chondrocyte group. The average wet mass of the low concentration residual ear chondrocytes in the control group was lower than that in the residual ear chondrocyte group (30%). (2) the average wet mass and the average content of glycosaminoglycan in co-transplantation group and residual ear chondrocyte group were significantly higher than those in the control group with high fat stem cells and low concentration of residual ear chondrocytes. Histological staining: mature cartilage lacunae were formed in co-transplantation group, residual ear chondrocyte group and low concentration residual ear chondrocyte control group. In low concentration residual ear chondrocytes, the cartilage lacunae were loosely arranged and the extracellular matrix was light in the control group. In the adipose stem cell group, fibroid tissue was found, and no cartilage lacunae were found in the adipose stem cell group. (4) Immunohistochemical staining of type 鈪，

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