Bals R, Gamarra F, Kaps A, Grundler S, Huber R M, Welsch U
Anatomische Anstalt, Chair II (Department of Cytology, Histology and Microscopical Anatomy), University of Munich, Munich, Germany.
Cell Tissue Res. 1998 Sep;293(3):573-7. doi: 10.1007/s004410051150.
To study the secretory products and the proliferation of cells of the human respiratory surface epithelium, we established a miniorgan-culture system of bronchial tissue. Biopsies of large airways were grown on agar-coated dishes immersed in a serum-enriched medium. As determined by light and transmission electron microscopy, between 1 and 3 weeks, the organ cultures were covered by a differentiated epithelium consisting of secretory, ciliated, and basal cells. Immunohistochemistry, using antibodies to mucin and lysozyme, and lectin histochemistry revealed both mucous and serous secretory cells in the epithelium. Cell proliferation was studied in situ using antibodies to proliferating cell nuclear antigen (PCNA) and Ki-67. Whereas at the time of explantation the proliferation was low (2.5+/-1.7% of the epithelial cells were PCNA-positive, 1.7+/-0.6 were Ki-67-positive), at 24 h of cultivation, 30.4+/-5.1% or 25.2+/-4.9% of the epithelial cells were labeled with antibodies to PCNA or Ki-67. After 7 days, the number of dividing cells was low again. The results show that the organ-culture system of human respiratory surface epithelium produces a differentiated epithelium that is useful in the study of secretory processes, differentiation, and proliferation.
为了研究人呼吸道表面上皮细胞的分泌产物和细胞增殖情况,我们建立了支气管组织的微型器官培养系统。将大气道活检组织置于浸有富含血清培养基的琼脂包被培养皿上培养。通过光镜和透射电镜观察确定,在1至3周内,器官培养物被由分泌细胞、纤毛细胞和基底细胞组成的分化上皮所覆盖。使用抗黏蛋白和溶菌酶抗体的免疫组织化学以及凝集素组织化学显示上皮中有黏液性和浆液性分泌细胞。使用抗增殖细胞核抗原(PCNA)和Ki-67抗体原位研究细胞增殖。在外植时增殖率较低(2.5±1.7%的上皮细胞PCNA阳性,1.7±0.6%的上皮细胞Ki-67阳性),而在培养24小时时,30.4±5.1%或25.2±4.9%的上皮细胞被PCNA或Ki-67抗体标记。7天后,分裂细胞数量再次减少。结果表明,人呼吸道表面上皮的器官培养系统产生了一种分化上皮,这对于研究分泌过程、分化和增殖是有用的。