Morphological characterization of the cells present in niches of the normal and reconstructed epithelia

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We attempted to produce novel monoclonal antibodies recognizing the niche cells of the human esophageal epithelium. Hybridomas were generated from fusions between spleen cells of ddY mice immunized with esophageal epithelial cells (EECs) cultured at low calcium concentrations. A clone, NJ-E-F4 was selected for its staining pattern on frozen sections of the epithelia, which produces a monoclonal antibody (moAb) reacting with almost cells of human esophageal epithelia, except the small populations of non-proliferating cells in the basal layer. The immunoreactivities of NJ-E-F4 were detected at Golgi area of cytoplasm in the cells predominantly located in the basal to the third layer. Ki67 positive proliferating cells ware observed in the second layer just above F4 negative cells in the basal layer, which were positive for our previous established NJ-E-H10 moAb. H10 moAb recognizes the basal cells in the interpapillary zones of the epithelia, and some of which seem to be the candidates of the stem cell in esophageal epithelia. The data seemed to be indicated that F4 negative, H10 positive and Ki67 negative cells are the candidates for the epithelial stem cells, which migrate into the second layer and start the cell cycle positive for Ki67. On the other hand, H10 positive cells in the rat skin were also observed in the hair follicle bulge which is the epidermis niche. So we used rat fetal H10 positive epithelia for the graft to grow the skin with hair follicles on the chorion of chick embryo. We succeeded to make the epidermis without differences of the H10 positive cells population and location between in the reconstructed and the normal. Therefore our moAb H10 and F4 are possibly to be good tools for the morphological examination of the epithelial niche cells.