Waldon D J, Kawabe T T, Baker C A, Johnson G A, Buhl A E
Upjohn Laboratories, Department of Dermatology Research, Kalamazoo, Michigan 49001.
In Vitro Cell Dev Biol Anim. 1993 Jul;29A(7):555-61. doi: 10.1007/BF02634149.
Inasmuch as hair follicles are difficult to maintain in culture, the study of hair biology using cultured hair follicles has met with only limited success. In our attempts to solve the problem of follicle degeneration, we cultured follicles at the air-surface interface on a modified collagen matrix (Gelfoam). In follicles cultured at the air-surface or submerged, we examined follicular morphology, hair shaft growth, sulfotransferase levels, cysteine incorporation, an expression of a tissue inhibitor of metalloproteinase (TIMP), and ultra-high sulfur keratin (UHSK). Follicles cultured at the air-liquid interface produced a 2.7-fold increase in hair growth and maintained an anagen-like morphology. Substrates such as nylon mesh seeded with fibroblasts, Full Thickness Skin, or 5-microns polycarbonate filter also supported hair growth, whereas Gelfilm, GF-A glass filter, filter paper, or 1-micron polycarbonate filter did not. The UHSK expression was significantly higher in the air-liquid interface cultures compared to the submerged culture. Several potassium channel openers, including minoxidil, a minoxidil analog, and the pinacidil analog (P-1075), all stimulated significant cysteine incorporation in follicles. Minoxidil and its analog specifically preserved the follicular root sheath, in contrast to P-1075 which did not, indicating a difference in the two drug types. The preservation of the root sheath was measured by increased TIMP expression and sulfotransferase activity and indicates that the root sheath is a target tissue for minoxidil. Our results show that follicles cultured at the air-liquid interface maintain a better morphology and produced greater hair growth than follicles cultured on tissue culture plastic.
由于毛囊在培养中难以维持，利用培养的毛囊进行毛发生物学研究仅取得了有限的成功。在我们试图解决毛囊退化问题的过程中，我们将毛囊培养在改良胶原基质（明胶海绵）的气液界面。对于在气液界面或浸没状态下培养的毛囊，我们检测了毛囊形态、毛干生长、磺基转移酶水平、半胱氨酸掺入、金属蛋白酶组织抑制剂（TIMP）的表达以及超高硫角蛋白（UHSK）。在气液界面培养的毛囊毛发生长增加了2.7倍，并维持了生长期样形态。接种成纤维细胞的尼龙网、全层皮肤或5微米聚碳酸酯滤膜等底物也支持毛发生长，而明胶膜、GF - A玻璃滤膜、滤纸或1微米聚碳酸酯滤膜则不支持。与浸没培养相比，气液界面培养中的UHSK表达显著更高。几种钾通道开放剂，包括米诺地尔、一种米诺地尔类似物和吡那地尔类似物（P - 1075），均刺激毛囊中半胱氨酸的显著掺入。米诺地尔及其类似物特异性地保留了毛囊根鞘，而P - 1075则没有，这表明两种药物类型存在差异。通过TIMP表达增加和磺基转移酶活性来衡量根鞘的保留情况，这表明根鞘是米诺地尔的靶组织。我们的结果表明，与在组织培养塑料上培养的毛囊相比，在气液界面培养的毛囊维持了更好的形态，并且毛发生长更多。
