Yeh T H, Herman P, Martin F, Soler P, Huy P T
Department of Oto-Rhino-Laryngology, Hopital Lariboisière, Paris, France.
Laryngoscope. 1993 Jun;103(6):645-52. doi: 10.1288/00005537-199306000-00012.
To overcome the technical difficulties encountered in investigating the physiology of Reissner's membrane (RM), primary culture of RM was established. Minute explants obtained by mechanical dissociation were seeded on impermeable substrate coated with collagen type I in serum-free, hormone-supplemented medium. Within 2 weeks, a large cultured area was obtained which reproduced the two cell layers of the original explant. One layer, deep or superficial depending on the side to which the explant adhered, was made of cells exhibiting light and electron microscopy characteristics of an epithelium. Immunohistochemical methods revealed that they expressed cytokeratin, demonstrating their epithelial nature. The second layer was made of mesenchymal cells. This study suggests that cell culture of RM will facilitate further investigation of its physiology.
为克服在研究赖斯纳膜（RM）生理学过程中遇到的技术难题，建立了RM原代培养体系。通过机械解离获得的微小外植体接种于涂有Ⅰ型胶原的不可渗透基质上，置于无血清、添加激素的培养基中。两周内，获得了一个大面积的培养区域，该区域重现了原始外植体的两层细胞。根据外植体附着的一侧，其中一层细胞位于深层或浅层，由具有上皮细胞光镜和电镜特征的细胞组成。免疫组织化学方法显示它们表达细胞角蛋白，证明了其上皮性质。第二层由间充质细胞组成。本研究表明，RM细胞培养将有助于进一步研究其生理学。
