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

正常和重建上皮细胞微环境中细胞的形态学特征

• 批准号: 14570006
• 负责人: HITOMI Jiro
• 金额: $ 2.62万
• 依托单位: Iwate Medical University (2003)Niigata University (2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570006/
• 关键词: epithelial niche; stem cells; monoclonal antibodies; immunohistochmistry; tissue engineering; skin; hair follicle; esophagus; 食道上皮; 肝細胞; 血管; 腺

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.

我们试图产生新的单克隆抗体识别的人食管上皮的小生境细胞。杂交瘤产生的ddY小鼠免疫与食管上皮细胞（EECs）在低钙浓度培养的脾细胞之间的融合。选择克隆NJ-E-F4，因为其在上皮的冷冻切片上的染色模式，其产生与人食管上皮的几乎所有细胞反应的单克隆抗体（moAb），除了基底层中的非增殖细胞的小群体。NJ-E-F4免疫反应阳性细胞主要分布于基底层至第三层细胞的胞质高尔基区。Ki 67阳性增殖细胞出现在第二层，位于基底层F4阴性细胞的正上方，这些细胞对我们先前建立的NJ-E-H10单克隆抗体呈阳性。H10单克隆抗体可识别食管上皮乳头间带的基底细胞，其中部分基底细胞可能是食管上皮干细胞的候选细胞。数据似乎表明，F4阴性、H10阳性和Ki 67阴性细胞是上皮干细胞的候选者，其迁移到第二层并开始Ki 67阳性的细胞周期。另一方面，大鼠皮肤中的H10阳性细胞也出现在毛囊隆突处，即表皮龛中。因此，我们采用大鼠胚胎H10阳性上皮细胞作为移植物，在鸡胚绒毛膜上长出带毛囊的皮肤。我们成功地使重建的表皮与正常表皮在H10阳性细胞的数量和位置上没有差异。因此，我们的单克隆抗体H10和F4可能是一个很好的工具，上皮龛细胞的形态学检查。

项目成果

Bcl11b is required for differentiation and survival of αβ T lymphocytes

• DOI: 10.1038/ni927
• 发表时间: 2003-06-01
• 期刊: NATURE IMMUNOLOGY
• 影响因子: 30.5
• 作者: Wakabayashi, Y;Watanabe, H;Kominami, R
• 通讯作者: Kominami, R

人見次郎, 平野茂樹: "ニワトリ胚漿尿膜上でラットの上皮組織を再構成する"解剖学雑誌. 78巻(抄録号). 199 (2003)

Jiro Hitomi、Shigeki Hirano：“鸡胚绒毛尿囊膜上的大鼠上皮组织的重建”解剖学杂志第 78 卷（摘要）。

上皮内niche(幹細胞成育微小環境)の形態学的解析と再構成系確立への試み

上皮内微环境（干细胞生长微环境）的形态分析并尝试建立重建系统

• 发表时间: 2005
• 作者: 人見 次郎

Bclllb is required for differentiation and survival of αβ T lymphocytes

Bclllb 是 αβ T 淋巴细胞分化和存活所必需的

• 发表时间: 2003
• 期刊: Nat Immunol 4
• 作者: Wakabayashi Y;Watanabe H;Inoue J;Takeda N;Sakata J;Mis hima Y;Hitomi J;Ymamamoto T;Utsuyama M;Niwa O;Aizaw a S;Kominami R.
• 通讯作者: Kominami R.

ニワトリ胚漿尿摸上でラットの上皮組織を再構成する

鸡胚尿囊绒毛膜大鼠上皮组织的重建

• 发表时间: 2003
• 期刊: 解剖学雑誌 78
• 作者: 人見 次郎;平野 茂樹
• 通讯作者: 平野 茂樹