Oliver A M
Immunopathology Laboratory, Department of Pathology, Aberdeen, Scotland, U.K.
Br J Dermatol. 1990 Dec;123(6):707-16. doi: 10.1111/j.1365-2133.1990.tb04187.x.
Skin samples taken from foetuses of 15-19 weeks gestational age and keratinocytes were cultured by the 3T3 feeder method or in serum-free MCDB 153 medium on 16 mm coverslips. Keratinocytes taken from paediatric circumcisions and patients undergoing plastic surgery were also cultured using the 3T3 feeder method. A panel of monoclonal antibodies against a number of cytokeratins and differentiation markers were used in the PAP technique to analyse the cells. Cryostat sections taken from the donor skin samples were stained simultaneously. Foetal skin expressed the cytokeratins 7, 13 and 19 that were not observed postnatally. This cytokeratin expression as maintained in both culture conditions and also observe in paediatric and adult keratinocytes. Cytokeratin 7 was expressed on a greater proportion of foetal cells than in vivo, whereas the expression of 13 and 19 decreased. All keratinocytes expressed vimentin, transferrin receptor and Ki67 (proliferating cell antigen), while a small proportion expressed involucrin throughout culture, indicating their high level of differentiation and proliferation. No differences were observed between low and high density cultures. Foetal keratinocytes cultured in MCDB 153 did not reach confluence and stronger staining of differentiation markers was observed in these cells. These results show that the in vivo differences in cytokeratin expression of foetal and adult keratinocytes disappear in the culture.
取自妊娠15 - 19周胎儿的皮肤样本和角质形成细胞,通过3T3饲养层法或在无血清的MCDB 153培养基中于16毫米盖玻片上进行培养。取自小儿包皮环切术和整形外科手术患者的角质形成细胞也采用3T3饲养层法进行培养。使用一组针对多种细胞角蛋白和分化标志物的单克隆抗体,采用PAP技术分析细胞。同时对取自供体皮肤样本的冷冻切片进行染色。胎儿皮肤表达细胞角蛋白7、13和19,这些在出生后未观察到。这种细胞角蛋白表达在两种培养条件下均得以维持,并且在小儿和成人角质形成细胞中也观察到。细胞角蛋白7在胎儿细胞中的表达比例高于体内,而13和19的表达则下降。所有角质形成细胞均表达波形蛋白、转铁蛋白受体和Ki67(增殖细胞抗原),而在整个培养过程中有一小部分表达兜甲蛋白,表明它们具有高度的分化和增殖水平。低密度和高密度培养之间未观察到差异。在MCDB 153中培养的胎儿角质形成细胞未达到汇合状态,并且在这些细胞中观察到分化标志物的染色更强。这些结果表明,胎儿和成人角质形成细胞在细胞角蛋白表达方面的体内差异在培养中消失。