| 李丹1 冯锐成1 张丽新1 郭方琪2 梁洋1 滕春波1.通过G418 处理离体纯化小鼠胰腺上皮细胞[J].,2011,11(12):2242-2246 |
| 通过G418 处理离体纯化小鼠胰腺上皮细胞 |
| Purification of Pancreatic Epithelial Cells Treated with G418 in Vitro |
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| DOI： |
| 中文关键词: G418 成纤维细胞 胰腺上皮细胞 |
| 英文关键词: G418 Fibroblasts Pancreatic epithelial cells |
| 基金项目:国家自然科学基金项目资助（30670304） |
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| 中文摘要: |
| 目的：探索一种能有效去除成纤维细胞，筛选有较强增殖能力上皮细胞的方法。方法：本研究利用成纤维细胞对G418 敏感
的特性，在成体小鼠胰腺细胞分离后，采用悬浮细胞直接接种到含30μg/mL G418 的培养基中，或细胞贴壁生长汇合至50-70%后
用30 至100μg/mL 之间不同浓度G418 处理24h、48h 或72h 两种方案进行上皮细胞的纯化。结果：在直接接种法培养处理中，
存活的大部分细胞为成纤维细胞，上皮细胞的生长受到抑制，无法得到纯化的胰腺上皮集落；而在细胞贴壁生长汇合至50-70%后
经G418 处理，成纤维细胞随着处理浓度的增加死亡率也在逐渐上升，其中50μg/mL G418 处理72 小时对去除成纤维细胞效果最
佳。结论：G418 处理能够有效去除成纤维细胞，分离纯化出一群在离体条件下具有强增殖能力、形成大上皮细胞集落的细胞。该
分离纯化方法为今后进一步研究成体胰腺干/ 祖细胞增殖与分化调控机制等问题奠定基础。 |
| 英文摘要: |
| Objective: To eliminate the fibroblasts and purify epithelial cells from adult mouse pancreas in vitro. Methods: Based
on the sensitivity of fibroblasts to G418, this study designed two schemes:Scheme 1 was that the isolated single cells from pancreas were
cultured directly in media with 30μg/mL G418, and Scheme 2 was that the isolated single cells were cultured to 50-70% aggregation, and
then they were treated with G418 at concentrations from 30 to 100 μg/mL of G418 for 24h, 48h or 72h. Results: The results showed that
the growth of the epithelial cells was inhibited in scheme 1, but that of the fibroblasts was little affected. However, after the pancreatic
cell grew 50-70% aggregation in scheme 2, G418 treatment could eliminate the fibroblasts, and the death ratio of fibroblasts increase with
the elevation of G418 concentration. Among them, 50 μg/mL G418 treatment for 72h was optimal to kill fibroblasts and preserve
epithelial cells. Conclutions: G418 treatment could effectively purify a pancreatic epithelial cells with strong proliferative and colonyforming
capability, thus provide a pathway for further studies on the proliferation and differentiation of adult pancreas stem/progenitor
cells. |
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