Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.
Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.
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在自发永生的非整倍性人角质形成细胞系中的正常角质化。
DOI：
10.1083/jcb.106.3.761
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发表时间：
1988-03
期刊：
影响因子：
--
通讯作者：
Fusenig NE
中科院分区：
文献类型：
--
作者：
Boukamp P;Petrussevska RT;Breitkreutz D;Hornung J;Markham A;Fusenig NE
与小鼠表皮细胞相比，人皮肤角质形成细胞对体外转化具有相当大的抗性。通过SV40已经实现了永生化，但是已经导致具有改变的分化的细胞系。我们已经建立了一个自发转化的人上皮细胞系，从成人皮肤，保持完整的表皮分化能力。该HaCaT细胞系显然是永生的（大于140代），在体外具有转化表型（在塑料和琼脂上的克隆形成），但保持非致瘤性。尽管HaCaT细胞具有改变的和无限的生长潜力，但与正常角质形成细胞相似，当移植到裸鼠上时，HaCaT细胞改革成有序结构和分化的表皮组织。分化特异性角蛋白（Nos. 1和10）和其他标记物（外皮蛋白和聚丝蛋白）表达并规则定位。因此，HaCaT是第一个来自成人皮肤的永久性上皮细胞系，其表现出正常分化，并为研究人细胞中角化的调节提供了有前途的工具。在核型分析中，该品系为非整倍体（最初为亚二倍体），具有独特的稳定标记染色体，表明单克隆起源。HaCaT系与起源组织的同一性通过使用高变小卫星探针的DNA指纹图谱来证明。这是第一次证明DNA指纹图谱不受长期培养、转化和多种染色体改变的影响，从而为明确鉴定人类细胞系提供了独特的可能性。HaCaT细胞系的特征清楚地证明，人成体角质形成细胞的自发转化可以在体外发生，并且与连续的染色体改变相关，尽管与分化中的主要缺陷没有必然联系。
In contrast to mouse epidermal cells, human skin keratinocytes are rather resistant to transformation in vitro. Immortalization has been achieved by SV40 but has resulted in cell lines with altered differentiation. We have established a spontaneously transformed human epithelial cell line from adult skin, which maintains full epidermal differentiation capacity. This HaCaT cell line is obviously immortal (greater than 140 passages), has a transformed phenotype in vitro (clonogenic on plastic and in agar) but remains nontumorigenic. Despite the altered and unlimited growth potential, HaCaT cells, similar to normal keratinocytes, reform an orderly structured and differentiated epidermal tissue when transplanted onto nude mice. Differentiation- specific keratins (Nos. 1 and 10) and other markers (involucrin and filaggrin) are expressed and regularly located. Thus, HaCaT is the first permanent epithelial cell line from adult human skin that exhibits normal differentiation and provides a promising tool for studying regulation of keratinization in human cells. On karyotyping this line is aneuploid (initially hypodiploid) with unique stable marker chromosomes indicating monoclonal origin. The identity of the HaCaT line with the tissue of origin was proven by DNA fingerprinting using hypervariable minisatellite probes. This is the first demonstration that the DNA fingerprint pattern is unaffected by long- term cultivation, transformation, and multiple chromosomal alterations, thereby offering a unique possibility for unequivocal identification of human cell lines. The characteristics of the HaCaT cell line clearly document that spontaneous transformation of human adult keratinocytes can occur in vitro and is associated with sequential chromosomal alterations, though not obligatorily linked to major defects in differentiation.