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Group 2 ILCs (ILC2s) produce cytokines such as IL\u20105 and IL\u201013 in response to alarmins, including IL\u201025 and IL\u201033 and play a key role in type 2 immunity. Lately, ILC2s have become a major focus across several research areas, including asthma, allergic lung inflammation, atopic dermatitis, and liver fibrosis. However, ILC2s are rare in human peripheral blood, making their isolation and expansion for functional studies challenging. Existing methods commonly rely on flow\u2010cytometry\u2010based cell sorting, which requires large sample volumes, expensive reagents, and considerable hands\u2010on time. Moreover, in vitro expansion protocols often depend on feeder cell lines and activating alarmins. Here, we present a simple and efficient approach for isolating and expanding human ILC2s from peripheral blood, without the need for flow\u2010cytometry\u2010based cell sorting, feeder cells, or external alarmins. This method yields pure and viable ILC2s from both large and small blood volumes, enabling the use of limited or rare patient samples. Expanded ILC2s responded robustly to alarmin stimulation by proliferating and producing signature cytokines and retained functionality after cryopreservation and reculture. Together, this method provides a user\u2010friendly and cost\u2010effective alternative to current human ILC2 isolation and expansion protocols.<\/jats:p>","DOI":"10.1155\/jimr\/7688318","type":"journal-article","created":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T06:34:47Z","timestamp":1787898887000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["An Optimized Feeder\u2010Free Method for Isolation and Expansion of Functional ILC2s Isolated From Human Peripheral Blood"],"prefix":"10.1155","volume":"2026","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-2685-4497","authenticated-orcid":false,"given":"Lina K.","family":"Andersson","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1639-8079","authenticated-orcid":false,"given":"Miriam","family":"Bollmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8534-9479","authenticated-orcid":false,"given":"Agnieszka","family":"Lastowska","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel","family":"Haupt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-7119-3369","authenticated-orcid":false,"given":"Lisi","family":"Wagener","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-4867-3254","authenticated-orcid":false,"given":"Anders","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4313-1521","authenticated-orcid":false,"given":"Johan","family":"Svensson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8931-0683","authenticated-orcid":false,"given":"Linda C.","family":"Johansson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8077-9824","authenticated-orcid":false,"given":"Mattias N. 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