Ex vivo development of a composite human skin equivalent from hair follicles micrografted

1. 我们从美国购买了AlloDerm，从韩国购买了SureDerm。我们决定在这个项目中使用“AlloDerm”，因为它在真皮结构和基底膜方面优于“AlloDerm”，我们对纤维连接蛋白、胶原蛋白IV和层粘连蛋白进行了HE和免疫组化染色。神户大学整形外科没有实验室，一开始我们得到了房间，并设置了大型设备，如洁净台，热灭菌器，冷冻离心机，培养箱，低温保存容器，工作台和固定的实验设施/仪器。我们于2003年初向神户大学伦理委员会申请资助培养表皮、口腔黏膜、毛囊细胞。我们在第二届日本再生医学年会上介绍了两种毛囊培养方法。由于“AlloDerm”的基膜成分中只存在层粘连蛋白，我们在“AlloDerm”中涂覆层粘连蛋白，并尝试进行毛囊移植，但均未成功。因此，我们尝试在其他基膜组件上涂覆，但后来都没有成功。目前，我们正在研究一种新的涂层基体预处理方法。另一方面，“AlloDerm”复合口腔黏膜的制造在神户大学取得成功，我们在临床应用中启动了伦理委员会，现在重复了一个案例（在第47届日本整形与重建外科学会年会上提出）。我们于2003年秋季购买气相色谱法，用三个月的时间完成仪器设置，分析毛囊和毛发的总脂肪酸。两者中都有大量的棕榈酸，而头发中不存在必需脂肪酸（在第三届日本再生医学年会和第十届瘢痕疙瘩/增生性疤痕会议上提出了这项研究）。此外，将于今年夏天在第二届世界伤口愈合学会上发表。现在我们正在对几个月前移植的口腔黏膜进行取样，并通过膜脂肪酸分析来检查它们是否在正常粘膜中恢复。少

1. We purchased "AlloDerm" from U.S.A. and "SureDerm" from Korea.2. We decided to utilize "AlloDerm" for this project because that was superior to this about dermal structure and basement membrane, performed HE and immunohistochemical staining of Fibronectin, Collagen IV, and Laminin.3. Kobe University plastic surgery did not have laboratory and at first we got the room and set large apparatus such as clean bench, heat sterilization apparatus, refrigerated centrifuge, incubator, cryopreservation vessel, bench and fixed experiment facilities/instrument.4. We applied to Kobe University ethic committee for allowance of cultivation epidermis, oral mucosa, hair follicle cells at the beginning of 2003.5. We presented two culture methods of hair follicles in the second Annual Meeting Japanese Regenerative Medicine.6. Because only Laminin was present among basement membrane components of "AlloDerm", we coated to "AlloDerm" in Laminin and tried hair follicle transp1antation, but did not succeed … More to cell inoculation even if take did it.7. So, we tried coating in other basement membrane components, but did not succeed afterwards. Now we are under study in attchin and a new coating matrix pretreatment.8. On the other hand, manufacture of the composite oral mucosa which used "AlloDerm" succeeds in Kobe University and we took off an ethic committee in clinical application and now repeats a case (presented it at the 47th Annual Meeting of Japan society plastic and Reconstructive surgery).9. We purchased gas chromatography in autumn, 2003, and instrumental setting completed it in three months and analyzed total fatty acids of hair follicles and hairs. Palmitic acid was a lot present in both, essential fatty acid was not present in hair (presented it in the third Annual Meeting of Japanese Regenerative Medicine, and the tenth keloid/hypertrophic scar meeting for the study. In addition, is to present it in 2nd World Wound Healing Society this summer).10. Now we are sampling the oral mucosa grafted several months ago, and to review whether they recover in normal mucosa through membranous fatty acid analysis. Less