兔舌黏膜细胞的体外培养及生物学特性研究

目的 探讨舌黏膜细胞的培养方法,为进一步以舌黏膜细胞作为种子细胞构建组织工程化角膜上皮提供实验依据.方法 体外切取舌侧面和底面黏膜组织,经0.25%Dispase分离上皮层后,再经0.05%的胰酶分离为单个细胞,接种在无血清角质细胞培养液(KSFM)中,进行体外培养,并对其形态学以及生长曲线、传代特点、各代生长特点进行观察.同时,对体外培养获得的舌黏膜细胞进行广谱角蛋白免疫荧光鉴定.结果 舌黏膜细胞能够在无血清角质细胞培养液中稳定生长,呈现典型的"铺路石"状,细胞形态单一,可传至3～4代.细胞免疫荧光显示,体外培养获得的舌黏膜细胞AE1/AE3免疫荧光染色阳性.结论 应用无血清角质细胞培养液,可在短期内获得大量具有增殖能力的舌黏膜细胞,为进一步将其作为种子细胞构建组织工程化角膜上皮奠定了良好的基础.

Experimental study on biological character of rabbit lingual keratinocyees

Objective To explore the cultural method of lingual keratinocytes and make preparations for further investigation in using lingual keratinocytes as a new choice of seed cells for the reconstruction of tissue engineered corneal epithelium.Methods Keratinocytes were enzymatically dissociated from lingual tissue specimens using 0.25%disepase and 0.05%trypsin.The biological characteristics of the cells were observed through phase microscope.Meanwhile.1ingual kemtinoeytes were immtmohistochemically labeled with broad spectrum keratin antibody (AE1/AF3).Results The cells revealed the form of "road metal" and could be passed for 3 or 4 generations.Immunohistochemistry indicated that lingual keratinocytes were positive for AE1/AE3 staining.Condusion Lingual kemtinocytes of rabbits can be cultured with KSFM in vitm and magnitude quantity can be attained,laying a favorable foundation for lingual keratinocytes as a new choice of seed cells for corneal epithelium reconstruction with tissue engineering.