Abstract
We present a protocol for isolation of putative epithelial progenitor cells from mouse lacrimal gland (LG) by fluorescence-activated cell sorting (FACS). Isolated LG epithelial progenitor cells can be cultured as 3D reaggregates within extracellular matrix gel or plated as a monolayer. 3D cultures could be maintained for several days and then dissociated with trypsin and plated as monolayer cultures, processed for analysis (e.g., mRNA/protein expression) and/or used for transplantations. Our goal is to provide researchers with a method that can be used as is or modified if isolation of other LG epithelial cell types is required.
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Acknowledgments
This protocol was adapted and modified from previous work published by both Helen Makarenkova and Robyn Meech and their respective research groups [4]. This work was supported by National Institutes of Health/National Eye Institute (NIH/NEI; Bethesda, MD, USA) Grants 1R01EY026202 (to HPM).
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Makarenkova, H.P., Meech, R. (2019). Isolation and Propagation of Lacrimal Gland Putative Epithelial Progenitor Cells. In: Bertoncello, I. (eds) Mouse Cell Culture. Methods in Molecular Biology, vol 1940. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9086-3_12
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DOI: https://doi.org/10.1007/978-1-4939-9086-3_12
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-9086-3
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