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从人类胃粘膜中分离密封环细胞的协议
Sarah G Samaranayake1, Lauren A Gamble1, Cassidy Bowden1
1Surgical Oncology Program, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
STAR protocols
|November 5, 2023
概括
生殖系致病性CDH1变体显著增加胃癌的风险. 这项研究引入了一种协议,用于从人类胃组织中分离可活的密封环细胞进行进一步分析.
科学领域:
- 在瘤学瘤学.
- 分子生物学分子生物学
- 胃肠病学 胃肠病学
背景情况:
- 生殖系致病性CDH1变体与患扩散型胃癌的风险>90%有关.
- 早期的,微观的封印环细胞癌通常是隐形的,但可以进展到晚期的疾病.
- 对这些早期细胞进行准确的表征对于了解疾病进展和制定治疗策略至关重要.
研究的目的:
- 提出一种用于从人类胃组织中获取密封环细胞的高活性悬浮的新方案.
- 为了促进下游分子和细胞分析早期的胃癌.
- 支持对CDH1相关胃癌的发病和早期检测的研究.
主要方法:
- 从人体组织样本中分离胃粘膜的详细方法.
- 逐步的组织解离技术,以实现单细胞悬浮.
- 细胞嵌入HistoGel以进行随后的免疫组织化学,流细胞测量和单细胞测序.
主要成果:
- 成功开发了一种产生高活性的密封环细胞悬浮物的协议.
- 证明该协议对免疫组织化学染色的实用性.
- 验证该协议的先进应用,包括流细胞计和单细胞测序.
结论:
- 本方案允许从人类胃组织中分离出可活的密封环细胞.
- 这种方法适用于各种下游分析,有助于研究早期胃癌.
- 该协议为研究人员研究CDH1相关的胃癌提供了宝贵的工具.
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