PROLIFERATIVE POTENTIAL OF DIFFERENT KERATINOCYTES OF PLUCKED HUMAN HAIR-FOLLICLES
PROLIFERATIVE POTENTIAL OF DIFFERENT KERATINOCYTES OF PLUCKED HUMAN HAIR-FOLLICLES
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DOI：
10.1111/1523-1747.ep12312406
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发表时间：
1995-07-01
影响因子：
6.5
通讯作者：
MOLL, I
中科院分区：
文献类型：
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作者：
MOLL, I
我们研究了集落形成能力，定位的集落形成细胞，并在体外寿命的外根鞘角质形成细胞的不同片段的成年人拔毛囊。细胞角蛋白和整合素的免疫组织化学染色显示这些细胞含有保存的基底细胞层。通过显微切割，分离出五个外根鞘片段（B1、B2、B3-1、B3-2、B4），用胰蛋白酶分散成单细胞悬液，并在人饲养层成纤维细胞上生长。所有片段都产生至少一些集落，但集落形成能力主要在中间部分（B2）和中央部分的下半部分（B3-1）中显著;毛囊的约60%的集落形成细胞定位于片段B3-1，28%定位于片段B3-2（中央部分的上半部分，包括凸起）。为了比较来自不同片段的细胞的体外寿命，我们在相同的条件下传代培养分离的角质形成细胞。条件片段B3-2中的片段长度最长，片段B1（鳞茎）中的片段长度最短。此外，通过对各种增殖和分化标记物的免疫细胞化学分析，研究了天然细胞和所有培养物的细胞在其整个寿命期间的分化状态。令人惊讶的是，角质形成细胞的所有片段，高分子量的细胞角蛋白和聚丝蛋白的表达所示，能够终末分化。这些数据表明，具有长寿命的细胞定位在靠近凸出区域的外根鞘的中央部分，并且具有高集落形成能力的细胞定位在中央下部。后者通常通过拔毛去除，因此可能不代表干细胞，而是在单个周期中对毛发生长重要的细胞。具有长寿命的细胞也包括在拔出的毛囊中可能是干细胞的直接后代，这些干细胞将被分离在隆突区域。最后，我们的研究结果对于遗传性皮肤病的基因转移和干细胞基因治疗是重要的，因为拔出的毛囊很容易获得，靠近凸起区域的角质形成细胞应选择性地使用。
We have examined colony-forming ability, localization of colony-forming cells, and in vitro life spans of outer root sheath keratinocytes of different fragments of adult human plucked hair follicles. These were shown by immunohistochemical staining for cytokeratins and integrins to contain a preserved basal cell layer. By microdissection, five fragments of the outer root sheath (B1, B2, B3-1, B3-2, B4) were separated, dispersed by trypsin into single cell suspensions, and grown on human feeder fibroblasts. All fragments gave rise to at least some colonies, but colony-forming ability was mostly marked in the intermediate part (B2) and the lower half of the central part (B3-1); approximately 60% of colony-forming cells of a hair follicle localized to the fragment B3-1 and 28% to the fragment B3-2 (upper half of the central part, including bulge). To compare the in vitro life spans of cells from the various fragments, we subcultured isolated keratinocytes under identical. conditions. The longest was found in the fragment B3-2 and the shortest in the fragment B1 (bulb). Moreover, the differentiation state of the native cells and the cells of all cultures were studied during their whole Life spans by immunocytochemical analysis of various proliferation and differentiation markers. Surprisingly, keratinocytes of all fragments, as shown by expression of high-molecular-weight cytokeratins and filaggrin, were capable of terminal differentiation. These data indicate that cells with long life spans are localized in central parts of the outer root sheath close to the bulge area and that cells with high colony-forming ability are localized in the lower central parts. The latter are usually removed by plucking and may therefore not represent stem cells but rather cells important for hair growth during a single cycle. Cells with long life spans-also included in plucked hair follicles-may be immediate progeny of stem cells that will be segregated in the bulge area. Finally, our results are important for gene transfer and stem cell gene therapy in genodermatoses, because plucked hair follicles are easily available and keratinocytes close to the bulge area should be used selectively.