Lanik Wyatt E, Xu Lily, Luke Cliff J, Hu Elise Z, Agrawal Pranjal, Liu Victoria S, Kumar Rajesh, Bolock Alexa M, Ma Congrong, Good Misty
Division of Newborn Medicine, Department of Pediatrics, Washington University School of Medicine.
Washington University.
J Vis Exp. 2018 Feb 15(132):56921. doi: 10.3791/56921.
Human small intestinal enteroids are derived from the crypts and when grown in a stem cell niche contain all of the epithelial cell types. The ability to establish human enteroid ex vivo culture systems are important to model intestinal pathophysiology and to study the particular cellular responses involved. In recent years, enteroids from mice and humans are being cultured, passaged, and banked away for future use in several laboratories across the world. This enteroid platform can be used to test the effects of various treatments and drugs and what effects are exerted on different cell types in the intestine. Here, a protocol for establishing primary stem cell-derived small intestinal enteroids derived from neonatal mice and premature human intestine is provided. Moreover, this enteroid culture system was utilized to test the effects of species-specific breast milk. Mouse breast milk can be obtained efficiently using a modified human breast pump and expressed mouse milk can then be used for further research experiments. We now demonstrate the effects of expressed mouse, human, and donor breast milk on the growth and proliferation of enteroids derived from neonatal mice or premature human small intestine.
人类小肠类器官源自隐窝,在干细胞生态位中生长时包含所有上皮细胞类型。建立人类类器官体外培养系统对于模拟肠道病理生理学以及研究相关特定细胞反应至关重要。近年来,来自小鼠和人类的类器官正在世界各地的多个实验室中进行培养、传代并保存以备将来使用。这个类器官平台可用于测试各种治疗方法和药物的效果以及它们对肠道中不同细胞类型产生的影响。在此,提供了一种从新生小鼠和早产人类肠道建立原代干细胞衍生的小肠类器官的方案。此外,利用这个类器官培养系统来测试物种特异性母乳的效果。使用改良的吸奶器可以高效获取小鼠母乳,然后将吸出的小鼠乳汁用于进一步的研究实验。我们现在展示了吸出的小鼠、人类和捐赠者母乳对源自新生小鼠或早产人类小肠的类器官生长和增殖的影响。