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Isolation and characterization of progenitor cells in uninjured, adult rat lacrimal gland

机译:成年大鼠泪腺未损伤祖细胞的分离和鉴定

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PURPOSE. The purpose of this study was to investigate the presence of progenitor cells in the uninjured, adult rat lacrimal gland (LG). METHODS. The presence of progenitor cells was examined in LG sections from male rats using antibodies against selected stem cell markers and α-smooth muscle actin (SMA), which marks myoepithelial cells (MECs), by immunofluorescence microscopy (IF). Small, immature cells were isolated after digestion of LG with collagenase and culture in RPMI 1640 for 2 weeks. Immature cells were examined for expression of stem cell markers by IF. Immature cell were grown in neuronal, epithelial, and myoepithelial cell media, and examined by light morphology and IF using antibodies to markers of different cell lineages. RESULTS. In the intact LGs, MECs expressed the stem cell markers nestin, Musashi 1, ABCG2, Pax6, Chx 10, ΔN p63, and Sox 2. All markers colocalized with SMA. Isolated immature cells contained Ki-67, nestin, Musashi 1, Pax 6, and CHX 10. In neuronal media, immature cells differentiated and assumed a neuronal cell morphology expressing neurofilament 200. In media for human corneal endothelial cells, immature cells differentiated, assumed cobblestone morphology, and labeled with the epithelial marker AE1/AE3. In RPMI media immature cells differentiated into cells with MEC-like morphology, and expressed the MEC markers SMA, α-actinin, adenylate cyclase II, and vimentin. CONCLUSIONS. We conclude that uninjured, adult LG contains progenitor cells that may be MECs, which can be isolated and differentiated into multiple lineages.
机译:目的。这项研究的目的是调查未受伤的成年大鼠泪腺(LG)中祖细胞的存在。方法。在雄性大鼠的LG切片中,使用针对所选干细胞标记物的抗体和标记平滑肌肌动蛋白(SMA)的抗体通过免疫荧光显微镜(IF)检测了雄性大鼠LG切片中祖细胞的存在。用胶原酶消化LG并在RPMI 1640中培养2周后,分离出未成熟的小细胞。通过IF检查未成熟细胞中干细胞标志物的表达。未成熟的细胞在神经元,上皮和肌上皮细胞培养基中生长,并使用针对不同细胞谱系标记的抗体通过光形态学和中频检查。结果。在完整的LG中,MEC表达干细胞标记Nestin,Musashi 1,ABCG2,Pax6,Chx 10,ΔNp63和Sox2。所有标记均与SMA共定位。分离出的未成熟细胞包含Ki-67,nestin,Musashi 1,Pax 6和CHX10。在神经元培养基中,未成熟细胞分化并呈现出表达神经丝200的神经元细胞形态。在人类角膜内皮细胞的培养基中,未成熟细胞得以分化。鹅卵石形态,并用上皮标记AE1 / AE3标记。在RPMI培养基中,未成熟细胞分化为具有MEC样形态的细胞,并表达MEC标记SMA,α-肌动蛋白,腺苷酸环化酶II和波形蛋白。结论。我们得出的结论是,未受伤的成年LG所包含的祖细胞可能是MEC,可以被分离并分化为多个谱系。

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