Long-term culture of mouse vibrissal dermal papilla cells and de novo hair follicle induction
Long-term culture of mouse vibrissal dermal papilla cells and de novo hair follicle induction
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DOI：
10.1089/ten.2006.0304
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发表时间：
2007-05-01
影响因子：
--
通讯作者：
Okochi, Hitoshi
中科院分区：
文献类型：
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作者：
Osada, Aki;Iwabuchi, Tokuro;Okochi, Hitoshi
我们已经成功地培养毛乳头（DP）细胞的长期和开发新的技术，提高他们的毛囊诱导效率在补丁分析。含成纤维细胞生长因子-2（FGF-2）的10%胎牛血清-Dulbecco改良基本培养基对小鼠触须DP细胞的生长有明显的刺激作用。此外，在连续培养（超过30代）期间保持增殖的效力。我们将这些已建立的DP细胞与表皮细胞结合，并将其皮下植入无胸腺小鼠，以检查其毛囊诱导能力。早期传代的DP细胞（第4代以下）可诱导新生毛囊，但后期传代的DP细胞不能诱导毛囊。接下来，我们将DP细胞聚集形成球体，然后将表皮细胞注射到它们中。与解离的DP细胞不同，由后期传代的细胞（超过10代）制成的球确实诱导新的毛囊。我们研究了几个特定的生长期卵泡DP基因，并证实，他们的表达水平升高的领域相比，他们在贴壁DP细胞的表达水平。这些结果表明，FGF-2是必不可少的毛乳头细胞培养和球的形成部分模型的完整DP，导致毛囊诱导，甚至后来传代细胞。
We have succeeded in culturing dermal papilla (DP) cells long term and developed new techniques that enhance their hair follicle-inducing efficiency in a patch assay. The outgrowing DP cells from mouse vibrissae were markedly stimulated by 10% fetal bovine serum-Dulbecco's modified essential medium that included fibroblast growth factor-2 (FGF-2). Moreover, the potency of proliferation was maintained during serial cultivations (more than 30 passages). We combined these established DP cells with epidermal cells and implanted them subcutaneously into athymic mice to examine their hair follicle-inducing ability. New hair follicles were induced by dissociated DP cells at earlier passages (under passage 4), but the cells from later passages could not induce follicles. We next aggregated the DP cells to form spheres and then injected them with epidermal cells. Unlike the dissociated DP cells, the spheres made from the later passaged cells (more than 10 passages) did induce new hair follicles. We examined several genes specific for DP of anagen follicles and confirmed that their expression level was elevated in the spheres compared with their expression level in adherent DP cells. These results suggest that FGF-2 is essential for dermal papilla cell culture and that sphere formation partially models the intact DP, resulting in hair follicle induction, even by later passaged cells.