Optimized method for higher yield of alveolar macrophage isolation for ex vivo studies.

Autor:	Devkota SP; Section of Microbiology and Immunology, Division of Biology, Kansas State University, Manhattan, KS, USA., Onah C; Section of Microbiology and Immunology, Division of Biology, Kansas State University, Manhattan, KS, USA., Joshi PR; Section of Microbiology and Immunology, Division of Biology, Kansas State University, Manhattan, KS, USA., Adhikari S; Section of Microbiology and Immunology, Division of Biology, Kansas State University, Manhattan, KS, USA., Baral P; Section of Microbiology and Immunology, Division of Biology, Kansas State University, Manhattan, KS, USA.
Jazyk:	angličtina
Zdroj:	Heliyon [Heliyon] 2024 Aug 30; Vol. 10 (17), pp. e37221. Date of Electronic Publication: 2024 Aug 30 (Print Publication: 2024).
DOI:	10.1016/j.heliyon.2024.e37221
Abstrakt:	Alveolar macrophages (AMs) are a fully differentiated lung-resident immune cell population and are a critical component of lung immunity. AMs can be easily isolated from mice via bronchoalveolar lavage fluid (BALF) collection. The quality and quantity of AMs in BALF isolation are critical for generating reliable and high-quality data for ex vivo studies. Traditional techniques use ice-cold (4°C) buffer to collect AMs in BALF and result in low yield. Hence, a new method that consistently gives a higher yield of AMs is needed. We demonstrate here an optimized method that significantly increases the quantity of AM recovery in BALF (>2.8 times than the traditional method). Our method uses a warm-buffer (37°C) containing EDTA. We compared the viability and functional parameters (cytokine/chemokine expression, phagocytosis) of AMs isolated by our new and traditional methods. Our study revealed that AMs collected using our method have similar viability and functional characteristics to those collected using traditional method. Hence, our new method can be used for the collection of a higher number of AMs without altering their function. This protocol might also be useful for isolating tissue-resident immune cells from other anatomical sites for ex vivo and other downstream applications. Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (© 2024 The Authors.)
Databáze:	MEDLINE
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