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通过流式细胞术从大鼠气管中分离和鉴定基底细胞和分泌细胞。

Separation and characterization of basal and secretory cells from the rat trachea by flow cytometry.

作者信息

Johnson N F, Wilson J S, Habbersett R, Thomassen D G, Shopp G M, Smith D M

机构信息

Inhalation Toxicology Research Institute, Lovelace Biomedical and Environmental Research Institute, Albuquerque, New Mexico 87185.

出版信息

Cytometry. 1990;11(3):395-405. doi: 10.1002/cyto.990110310.

Abstract

Basal and secretory cells have been separated as highly enriched viable populations from single-cell suspensions of rat tracheal epithelial cells. Isolation of the populations was achieved by preparation of a cell suspension and separation by flow cytometry using contour maps generated from 2 degrees and 90 degrees light scatter signals. Flow cytometric analysis of cells showed 10% of the whole preparation were cells in SG2M phase of the cell cycle. The secretory cells accounted for 86% of these cycling cells; the remainder were accounted for by the basal cells. Culture of sorted populations of basal and secretory cells in serum free defined medium showed that basal cells had a lower (0.6%) colony-forming efficiency than secretory cells (3.4%). Significant differences in blue auto-fluorescence, Hoechst 33342 uptake, and lectin staining were apparent between basal and secretory cells. These results suggest that the secretory cell rather than the basal cell is primarily the cell type involved in maintenance of the normal tracheal epithelium. Secretory cells are greater in number, have a higher proliferative potential, and greater metabolic capability. Because of these traits they may be a critical cell at risk from damage by environmental agents.

摘要

已从大鼠气管上皮细胞的单细胞悬液中分离出基底细胞和分泌细胞,作为高度富集的活细胞群体。通过制备细胞悬液并利用由2°和90°光散射信号生成的等高线图进行流式细胞术分离,实现了这些细胞群体的分离。细胞的流式细胞术分析显示,整个制备物中有10%的细胞处于细胞周期的SG2M期。分泌细胞占这些循环细胞的86%;其余的由基底细胞构成。在无血清限定培养基中培养分选后的基底细胞和分泌细胞群体,结果显示基底细胞的集落形成效率(0.6%)低于分泌细胞(3.4%)。基底细胞和分泌细胞在蓝色自发荧光、Hoechst 33342摄取和凝集素染色方面存在明显差异。这些结果表明,分泌细胞而非基底细胞主要是参与维持正常气管上皮的细胞类型。分泌细胞数量更多,具有更高的增殖潜能和更强的代谢能力。由于这些特性,它们可能是易受环境因素损伤的关键细胞。

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