DIFFERENTIATION SPECIFIC FUNCTIONS IN CULTURED AND TRANSPLANTED MOUSE KERATINOCYTES - ENVIRONMENTAL-INFLUENCES ON ULTRASTRUCTURE AND KERATIN EXPRESSION
DIFFERENTIATION SPECIFIC FUNCTIONS IN CULTURED AND TRANSPLANTED MOUSE KERATINOCYTES - ENVIRONMENTAL-INFLUENCES ON ULTRASTRUCTURE AND KERATIN EXPRESSION
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DOI：
10.1111/j.1432-0436.1984.tb01389.x
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
FUSENIG, NE
中科院分区：
文献类型：
--
作者：
BREITKREUTZ, D;BOHNERT, A;FUSENIG, NE
从新生小鼠背部皮肤中提取的角质形成细胞，在常规条件下（浸泡）在原代培养（PEC）中生长，从细胞形态和组织、超微结构和角蛋白组成来看，其分化程度有所降低。在初代细胞骨架提取物中发现了四种主要的角蛋白（分子量为60k， 59K, 53K, 49K），以及少量的51K蛋白和一些残留的肌动蛋白。这种培养型角蛋白谱在不同药物（如12-0-十四烷醇-13-乙酸酯（TPA）、维甲酸（RA）或二甲基亚砜（DMSO））反复处理后保持稳定，几乎没有变化。原代培养物在低Ca2+ (0.1 mM)培养基或3T3喂料层中长期生长，其性状没有改变。然而，TPA、RA和低Ca2+确实改变了形态学和纤维结构（通过间接免疫荧光显微镜观察到抗角蛋白抗体）。将原代培养的角质形成细胞与从表皮分离的基底细胞进行比较，发现两者在电泳图谱上具有相似性，其中60K和53K角化蛋白是共同的。其他体内角蛋白具有特征性的空间分布；67K和58K角蛋白存在于基底上细胞（棘状和粒状），而64K、62K、58.5K和57.5K角蛋白仅存在于角质层。在常规条件下培养的细胞中没有发现这些角蛋白。PEC生长在暴露于空气（器官型培养）的胶原凝胶上，可以恢复表皮分化的几种形态特征，而不影响角蛋白谱。PEC移植到成年同基因小鼠身上后，表皮功能几乎完全恢复。在这种体内环境中，发育了结构良好的上皮，类似于滤泡间表皮。典型超微结构和体内型角蛋白表达均在2或3周内恢复。虽然原代培养的角质形成细胞与体内的有很大的不同，但它们并没有不可逆地失去完全分化的能力。
Keratinocytes from neonatal mouse back skin, growing in primary culture (PEC) under conventional conditions (immersed), exerted a reduced program of differentiation as indicated by cell morphology and organization, ultrastructure and keratin composition. Four major keratins were found in cytoskeletal extracts (MW 60 K [kilodalton], 59K, 53K, 49K) of primary cltures, together with a minor 51K protein and some residual actin. This culture-type keratin profile remained stable and little variation was observed after repeated treatment with various agents, such as 12-0-tetradecanoylphorbol-13-acetate (TPA), retinoic acid (RA) or dimethylsulfoxide (DMSO). The profile was unaltered by long-term growth of primary cultures in low Ca2+ (0.1 mM) medium or on 3T3 feeder-layers. Nevertheless, TPA, RA and low Ca2+ did alter the morphology and filament architecture (as observed by indirect immunofluorescence microscopy with anti-keratin antibodies). Comparison of keratinocytes from primary culture with basal cells isolated from epidermis revealed similarities in electrophoretic profile, with the 60K and 53K keratins being common to both. The other in vivo keratins had a characteristic spatial distribution; 67K and 58K keratins were present in suprabasal cells (spinous and granular), while 64K, 62K, 58.5K and 57.5K keratins were present only in stratum corneum. None of these keratins were found in cultured cells grown under regular conditions. Several morphological features of epidermal differentiation could be restored by the growth of PEC on collagen gels exposed to the atmosphere (organotypic culture) without influencing the keratin profile. Almost complete restoration of epidermal function was achieved after transplantation of PEC onto adult syngeneic mice. In this in vivo environment, well-structured epithelia developed which resembled interfollicular epidermis. Restoration of both typical ultrastructure and in vivo type keratin expression occurred within 2 or 3 wk. Although keratinocytes in primary culture differ considerably from those in vivo, they have not irreversibly lost the capacity for complete differentiation.