一种简单而快速的方法,同时隔离小鼠的初级岛屿和初级胰腺状细胞
Xiaorong Tian1, Hanxiao Cui2, Jiayu Li2
1Clinical Medical College, Yangzhou University; Department of Gastroenterology, Shanghai Institute of Pancreatic Diseases, Changhai Hospital, National Key Laboratory of Immunity and Inflammation, Naval Medical University.
Journal of visualized experiments : JoVE
|January 26, 2026
概括
研究人员开发了一种更简单的方法来隔离主要胰腺细胞,用于研究胰腺炎后的糖尿病. 这种技术避免了复杂的体内输液,产生高质量的细胞用于研究胰腺外分泌内分泌相互作用.
科学领域:
- 内分泌学和新陈代谢学
- 细胞生物学 细胞生物学
- 手术技巧 手术技巧
背景情况:
- 研究后急性胰腺炎糖尿病 (PPDM-A) 需要了解胰腺细胞 (PAC) 和小岛细胞通信.
- 现有的分离初级小岛的方法涉及技术上具有挑战性的体内 perfusion,限制了许多研究人员的可访问性.
- 不朽化的细胞系不能准确地复制体内病理生理条件,需要高质量的初级细胞隔离.
研究的目的:
- 开发和验证一种简化的体外法,用于从小鼠中分离初级胰岛和细胞.
- 为获得可靠的胰腺细胞提供可靠的技术,用于研究外分泌内分泌相互作用和PPDM-A病原性.
主要方法:
- 建立了一个修改后的协议,消除了对活体胰腺输液的需要.
- 从被安乐死的SPF级C57BL/6J小鼠中分离胰腺,然后用原酶P进行体外消化.
- 通过Ficoll密度梯度离心分离初级小岛和通过过和离心分离的acinar细胞.
- 使用卡尔/PI染色,葡萄糖刺激胰岛素分泌和氨酶活性测定来评估细胞活力和功能.
主要成果:
- 经过修改的方法证明了操作的方便性和高细胞产量:每只小鼠有 (120 ± 5) 的小岛和 (1.61.95 × 107) 的小岛细胞.
- 对于孤立的小岛群 ((97.52 ± 0.16)%) 和小岛细胞 ((96.55 ± 0.95)%) 实现了高生存率.
- 孤立的小岛保持了正常的胰岛素分泌能力,状细胞对状蛋白刺激作出了有效的反应.
结论:
- 简化的体外试验方法提供了一种可靠且易于使用的方法来隔离初级胰岛和细胞.
- 这种技术有助于研究胰腺内分泌外分泌相互作用和PPDM-A.A.的致病性.
- 该方法对大鼠的适用性仍需得到验证.
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