Hammar H, Acevedo F, Naito S
Department of Dermatology, Karolinska Hospital, Karolinska Institute, Stockholm, Sweden.
Acta Derm Venereol. 1990;70(1):11-7. doi: 10.2340/00015555701117.
Growth of keratinocytes in explant culture of mouse ear epidermis was studied. The addition of transferrin to the culture media improved growth. Transferrin fractionated from human and fetal calf serum increased outgrowths of the cultures when compared with commercially available transferrin. An acidic transferrin fraction was present in greater amount in human serum and in fetal calf serum than that found in commercial transferrin. This fraction was more abundant in serum from psoriatic patients than in serum of healthy subjects as shown by isotachophoresis. For the culture studies, preparation of this material was done by chromatography on DEAE-Sepharose 6B-CL columns. Further on, diferric transferrin was preferentially used in order to abolish variation due to iron saturation. Iron concentration higher than 5 microM was deleterious to cell growth. The basal culture medium contained transferrin depleted fetal calf serum in RPMI 1640 with 2 microM glutamine and antibiotics. Serum-free medium was used in some experiments. The additions were 1.7 microM insulin, 1.4 microM hydrocortisone, 10 microM ethanolamine and 10 microM phosphoethanolamine. A partially purified fraction of the acidic forms of transferrin (10-20 micrograms/ml medium) improved outgrowth when compared with a neutral fraction under these circumstances.
研究了小鼠耳表皮外植体培养中角质形成细胞的生长情况。向培养基中添加转铁蛋白可促进生长。与市售转铁蛋白相比，从人血清和胎牛血清中分离得到的转铁蛋白能增加培养物的生长。人血清和胎牛血清中的酸性转铁蛋白组分比市售转铁蛋白中的含量更高。等速电泳显示，银屑病患者血清中的该组分比健康受试者血清中的更丰富。在培养研究中，通过在DEAE-琼脂糖6B-CL柱上进行色谱法制备该物质。此外，优先使用二价铁转铁蛋白以消除因铁饱和度引起的差异。铁浓度高于5 microM对细胞生长有害。基础培养基在RPMI 1640中含有去除转铁蛋白的胎牛血清、2 microM谷氨酰胺和抗生素。在一些实验中使用了无血清培养基。添加物包括1.7 microM胰岛素、1.4 microM氢化可的松、10 microM乙醇胺和10 microM磷酸乙醇胺。在这些情况下，与中性组分相比，部分纯化的酸性形式转铁蛋白组分（10 - 20微克/毫升培养基）能改善生长。
