Frech Michael, Knipfer Lisa, Wirtz Stefan, Zaiss Mario M
Department of Internal Medicine 3, Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg (FAU) and Universitätsklinikum Erlangen, Erlangen, Germany.
Deutsches Zentrum für Immuntherapie (DZI), Germany.
MethodsX. 2020 Sep 10;7:101054. doi: 10.1016/j.mex.2020.101054. eCollection 2020.
Group 2 innate lymphoid cells (ILC2s) are a recently recognized subset of innate lymphocytes with crucial role in mucosal immunity and tissue homeostasis. Over the past decade, substantial advances in our understanding of ILC2 biology have established them as an essential element in innate and adaptive immunity. However, their relatively low abundance and laborious purification from mucosal tissues make their study difficult. Moreover, due to a lack of an ILC2-specific Cre mouse-line, adoptive transfer of ILC2s into ILC-deficient hosts is inevitable. Herein we describe an in-depth protocol for the induction, isolation, and expansion of murine ILC2s. By combining an in vivo gene delivery approach to boost ILC2 numbers and a cell culture strategy to expand isolated cells, large quantities of highly pure ILC2s can be obtained. The isolated cells maintain their phenotype and can be used for subsequent cell transfer or in vitro studies. In comparison to previous protocols, this approach is cost-effective and efficient with potential yield of more than 20 million ILC2s isolated per mouse. • • •
第2组固有淋巴细胞（ILC2s）是最近才被认识的固有淋巴细胞亚群，在黏膜免疫和组织稳态中起关键作用。在过去十年里，我们对ILC2生物学特性的理解取得了重大进展，确立了它们在固有免疫和适应性免疫中的重要地位。然而，它们在黏膜组织中的丰度相对较低，且从黏膜组织中纯化的过程繁琐，这使得对它们的研究颇具难度。此外，由于缺乏ILC2特异性的Cre小鼠品系，将ILC2过继转移到ILC缺陷宿主中是不可避免的。在此，我们描述一种用于诱导、分离和扩增小鼠ILC2s的详细方案。通过结合体内基因递送方法以增加ILC2数量和细胞培养策略来扩增分离的细胞，可以获得大量高纯度的ILC2s。分离出的细胞保持其表型，可用于后续的细胞转移或体外研究。与之前的方案相比，该方法具有成本效益且高效，每只小鼠潜在的ILC2s分离产量超过2000万个。 • • •
