Abstract
Intestinal fibroblasts maintain homeostasis and contribute to inflammatory responses and the development of cancer. Intestinal fibroblasts express pattern recognition receptors which can mount an immune response. Since intestinal fibroblasts interact with diverse immune and nonimmune cells, further insights into the biology of intestinal fibroblasts could expand our knowledge of the development, homeostasis, and pathophysiology of the intestine. Here, we describe a simple protocol for the isolation, cultivation, and maintenance of primary fibroblasts from the mouse colon. These cells express α-smooth muscle actin, a characteristic of specialized contractile fibroblasts called myofibroblasts. We also outline the use of these colonic fibroblasts for immunoblotting and immunofluorescence assays with or without stimulation with a growth factor.
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Acknowledgements
The authors acknowledge the facilities and the scientific and technical assistance of Microscopy Australia at the Centre for Advanced Microscopy (ANU, Australia), a facility that is funded by the University and the Federal Government. The authors acknowledge the National Collaborative Research Infrastructure Strategy (NCRIS) via Phenomics Australia. A.P. and M.K. are supported by the Gretel and Gordon Bootes Medical Research Foundation. M.K. is supported by the John Curtin School of Medical Research PhD Scholarship. S.M.M. is supported by the Australian National University, the National Health and Medical Research Council of Australia (under Project and Ideas Grants APP1146864 and APP2002686), and a CSL Centenary Fellowship.
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Pandey, A., Kurera, M., Man, S.M. (2023). Primary Intestinal Fibroblasts: Isolation, Cultivation, and Maintenance. In: Jenkins, B.J. (eds) Inflammation and Cancer. Methods in Molecular Biology, vol 2691. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3331-1_23
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DOI: https://doi.org/10.1007/978-1-0716-3331-1_23
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3330-4
Online ISBN: 978-1-0716-3331-1
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