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

我们试图制备识别人食管上皮小生境细胞的新型单克隆抗体。食道上皮细胞（EECs）经低钙培养免疫后，小鼠脾脏细胞融合形成杂交瘤。选择一个克隆NJ-E-F4，其在上皮冷冻切片上的染色模式，产生一种单克隆抗体（moAb），与几乎所有人食管上皮细胞反应，除了基底层的少量非增殖细胞。NJ-E-F4在以基底至第三层细胞为主的细胞质高尔基区检测到免疫反应。在基底层F4阴性细胞上方的第二层观察到Ki67阳性增殖细胞，对我们先前建立的NJ-E-H10 moAb呈阳性。H10 moAb识别上皮乳头间带的基底细胞，其中一些似乎是食管上皮干细胞的候选细胞。这些数据似乎表明，F4阴性、H10阳性和Ki67阴性的细胞是上皮干细胞的候选者，它们迁移到第二层并开始Ki67阳性的细胞周期。另一方面，在大鼠皮肤表皮生态位毛囊膨出处也观察到H10阳性细胞。因此，我们用大鼠胎H10阳性上皮细胞移植到鸡胚绒毛膜上生长毛囊皮肤。我们成功地使重建的表皮与正常的表皮没有H10阳性细胞的数量和位置的差异。因此，我们的moabeh10和F4可能是对上皮小生境细胞进行形态学检查的良好工具。

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.