發(fā)布時(shí)間：2018-02-03 00:49

  本文關(guān)鍵詞： 軟骨細(xì)胞 脂肪類 干細(xì)胞 組織工程 脂肪干細(xì)胞 殘耳軟骨細(xì)胞 脂肪來源干細(xì)胞 軟骨形成 共移植 誘導(dǎo) 體內(nèi)　出處：《中國組織工程研究》2017年21期 　論文類型：期刊論文

【摘要】：背景:先天性小耳畸形殘耳軟骨細(xì)胞從細(xì)胞數(shù)量和質(zhì)量上難以作為種子細(xì)胞構(gòu)建出正常人耳郭大小的同質(zhì)耳軟骨支架。目的:探討殘耳軟骨細(xì)胞能否在裸鼠體內(nèi)非軟骨環(huán)境下模擬軟骨誘導(dǎo)微環(huán)境,促進(jìn)脂肪來源的干細(xì)胞向軟骨分化并形成組織工程軟骨,從而為解決組織工程化人耳郭軟骨支架制備做基礎(chǔ)準(zhǔn)備工作。方法:(1)實(shí)驗(yàn)分為4組:第2代殘耳軟骨細(xì)胞與第3代脂肪來源的干細(xì)胞以3∶7比例混合作為共移植組,單純殘耳軟骨細(xì)胞作為陽性對(duì)照組(殘耳軟骨細(xì)胞組),單純脂肪來源的干細(xì)胞作為陰性對(duì)照組(脂肪干細(xì)胞組),以上3組接種細(xì)胞終濃度為5.0×10~(10) L~(-1),低濃度殘耳軟骨細(xì)胞對(duì)照組細(xì)胞終濃度為1.5×10~(10) L~(-1);(2)按照實(shí)驗(yàn)分組將0.2 mL細(xì)胞-Pluronic-F127復(fù)合物注射到裸鼠背部皮下,體內(nèi)培養(yǎng)8周后對(duì)新生組織進(jìn)行大體觀察、濕質(zhì)量測量、糖胺多糖含量測定、組織學(xué)及免疫組化染色檢測。結(jié)果與結(jié)論:(1)共移植組平均濕質(zhì)量達(dá)到殘耳軟骨細(xì)胞組的80%以上;低濃度殘耳軟骨細(xì)胞對(duì)照組平均濕質(zhì)量低于殘耳軟骨細(xì)胞組的30%;(2)共移植組和殘耳軟骨細(xì)胞組的平均濕質(zhì)量和糖胺多糖平均含量均顯著高脂肪干細(xì)胞組和低濃度殘耳軟骨細(xì)胞對(duì)照組(P0.05);(3)組織學(xué)染色:共移植組、殘耳軟骨細(xì)胞組與低濃度殘耳軟骨細(xì)胞對(duì)照組標(biāo)本均有成熟的軟骨陷窩形成,低濃度殘耳軟骨細(xì)胞對(duì)照組軟骨陷窩松散排列不均,胞外基質(zhì)著色淡;脂肪干細(xì)胞組為纖維樣組織,未見軟骨陷窩形成;(4)Ⅱ型膠原免疫組化染色:共移植組、殘耳軟骨細(xì)胞組與低濃度殘耳軟骨細(xì)胞對(duì)照組可見成熟軟骨陷窩周圍有不同程度的棕黃色沉淀即Ⅱ型膠原表達(dá);脂肪干細(xì)胞組未見Ⅱ型膠原表達(dá);(5)結(jié)果提示,殘耳軟骨細(xì)胞能夠在裸鼠體內(nèi)非軟骨環(huán)境下模擬軟骨誘導(dǎo)微環(huán)境,促進(jìn)脂肪干細(xì)胞向軟骨分化并生成組織工程軟骨。
[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，

本文編號(hào)：1485938