CAJ | 학술논문

采集婴儿皮肤组织,用组织块法制备原代细胞,通过差速消化和差速贴壁法纯化细胞,扩大培养至第3代液氮冻存,复苏细胞后进行活力、形态、生长曲线、微生物污染、核型、乳酸脱氢同工酶谱及荧光蛋白质粒转染表达等特性研究.结果显示,婴儿皮肤组织细胞呈成纤维型,生长状态良好；生长曲线呈“S”型,最大增值浓度为1.64×105/mL,倍增时间为63.12 h；细菌、真菌、支原体检测均为阴性；染色体2n=46,二倍体为86%,占主体；乳酸脱氢酶同工酶电泳图谱显示有5条谱带；外源质粒在该细胞中能进行复制和表达.表明已成功建立了婴儿皮肤组织成纤维细胞系,这为复合人工皮肤的组织工程学研究奠定了前期工作基础,也为皮肤相关疾病的研究提供了实验条件,因而具有重要的理论和实践意义.
채집영인피부조직,용조직괴법제비원대세포,통과차속소화화차속첩벽법순화세포,확대배양지제3대액담동존,복소세포후진행활력、형태、생장곡선、미생물오염、핵형、유산탈경동공매보급형광단백질립전염표체등특성연구.결과현시,영인피부조직세포정성섬유형,생장상태량호；생장곡선정“S”형,최대증치농도위1.64×105/mL,배증시간위63.12 h；세균、진균、지원체검측균위음성；염색체2n=46,이배체위86%,점주체；유산탈경매동공매전영도보현시유5조보대；외원질립재해세포중능진행복제화표체.표명이성공건립료영인피부조직성섬유세포계,저위복합인공피부적조직공정학연구전정료전기공작기출,야위피부상관질병적연구제공료실험조건,인이구유중요적이론화실천의의.
The baby skin explants were cultured with tissue pieces method. The primary cells were purified and preserved with liquid nitrogen after being passaged to the thrid generation. Observations on morphology,dynamic growth,microbial contamination, analysis of karyotype, isoenzyme of lactate dehydrogenase and fluorescin plasmid transfection and expression were carried out. The results showed that the cell line was fibroblast and growed well. The maximum proliferation concentration was 1. 64×105/mL and poulation doubling time of cells(PDT)was 63. 12 h. The tests for bacteria,fungi,virus and mycoplasma were all negative. The chromosomes were 46 and diploid cells were dominant of 86%. The banding patterns of the isoenzyme of lactate dehydrogenase had significant difference and there were five bands in all. The exogenous plasmid could copy and express in the cell. Those showed that the cell line was established successfully,which lays the foundation of the work bases for composite artificial skin tissue engineering and provides the experimental conditions for skin diseases. So this has the important theoretical and practical significance.