摘要
背景:耻骨联合是与关节盘、半月板或椎间盘具有相同性质的组织,但是将其作为组织工程种子细胞供区的研究,至今鲜有报道。目的:观察耻骨联合作为组织工程种子细胞供区的可行性。方法:取大鼠的耻骨联合组织进行苏木精-伊红、阿尔新蓝及Ⅱ型胶原免疫组织化学染色,观察细胞外基质的组织学特征。再将分离自大鼠耻骨联合组织的细胞,在体外培养扩增后,与藻酸盐凝胶支架复合立体培养,通过活细胞荧光标记方法,观察细胞存活率,通过21d长期培养观察细胞增殖情况。测量8具成年男性骨盆标本的耻骨联合软骨区的体积,判断其是否有利于分离足够数量的种子细胞。结果与结论:苏木精-伊红染色显示,大鼠耻骨联合组织有大量平行或交叉排列的纤维束,细胞单独散在、成对或成行排列分布于纤维束之间的陷窝内。阿尔新蓝染色和Ⅱ型胶原免疫组织化学染色发现,大鼠耻骨联合组织广泛着色,细胞及陷窝附近着色较深。藻酸盐凝胶支架为多孔结构,孔隙间交互通连,孔隙直径为(27.0±16.7)μm,孔隙壁较光滑。在支架上细胞存活率为(72.4±4.5)%。在体外长期培养过程中,支架上的细胞呈圆球形,逐渐形成"同源细胞群",培养13d时观察到的细胞团,在第21天时明显增大,所含细胞数明显增多。成年男性耻骨联合组织的体积为(1.13±0.21)cm3。结果证实,大鼠耻骨联合组织细胞具有软骨细胞的排列特征,细胞外间质有软骨组织特异性基质成分。取自大鼠耻骨联合的细胞,在体外立体培养具有持续的增殖能力,耻骨联合组织可作为软骨组织工程种子细胞供区。成年男性耻骨联合组织量较大,有利于分离获取较多的原代细胞。
BACKGROUND： Pubic symphysis is the same kind of tissue as the temporemandibular joint disc, meniscus or intervertebral disc, but it is rarely reported that pubic symphysis can be used as a donor area of tissue engineering seed cells. OBJECTIVE： To explore the feasibility of the pubic symphysis as the donor area of tissue engineering seed ceils. METHODS： Hematoxylin-eosin stain, alcian blue stain and type Ⅱ collagen immunohistochemical study were used to detect the histological features of the extracellular matrix of rat pubic symphysis tissue. Cells derived from rat pubic symphysis were cultured in three-dimensional alginate scaffold in vitro. Cell viability in scaffold was detected by living cells fluorescence staining. Cells proliferation in the alginate scaffold cultured in vitro for 21 days was observed. Eight pelvis specimens of adult men were used to measure the cartilage volume of the pubic symphysis. RESULTS AND CONCLUSION： Parallel and crossing fibrous bands were spread in the rat pubic symphysis tissue, and between them there were small round or short-fusiform cells in the lacunae, with basophilic cytoplasm. Cells were in pairs or lined up. Sections stained with alcian blue and type Ⅱ collagen antibody showed positive in the cytoplasm and extracellular matrix surrounding the round lacunae, whereas the matrix farther from cells stained lightly. Alginate gel scaffold displayed a porous structure with （27.0±16.7） μm pore diameter. Cell viability in the scaffold was （72.4±4.5）%. The spheroidal cells in the alginate gel grew into isogenous group after long-term three-dimensional culture in vitro. Cell clusters were observed after cultured for 13 days, and became larger with more cells after 21 days culture. Pubic symphysis cartilage region of adult man was wedge shaped with thick front part and its volume was （1.13±0.21） cm2. It turns out that cells of the rat pubic symphysis have the chondrocyte features as alignment and specific histological stain of cartilage tissue. Cells derived from the pubic symphysis proliferate well in three-dimensional culture in vitro. Volume of adult man pubic symphysis cartilage is large enough as a donor site to supply cartilage mass for cells isolation.
出处
《中国组织工程研究》
CAS
CSCD
2013年第11期1972-1980,共9页
Chinese Journal of Tissue Engineering Research
基金
广东省科学技术厅基金资助项目(2004A30201004)~~
关键词
组织构建
骨组织构建
耻骨联合
形态学
软骨
细胞外基质
增殖
立体培养
种子细胞供区
组织工程
省级基金
组织构建图片文章
tissue construction
bone tissue construction
pubic symphysis
morphology
cartilage
extracellular matrix
proliferation
three-dimensional culture
donor area of seed cells
tissue engineering
provincial grants-supported paper
tissue construction photographs-containing paper
