I want to isolate tracheal epithelial cells from mice. To deplete fibroblasts from the primary culture, usually Primaria BD plates are used. Can anybody suggest some other substitute or which can prepared in lab? Thanks
Hi Bansal
Look at abstract of Lam et al 2011 and PDF attached i think help you
J Vis Exp. 2011; (48): 2513.
Published online 2011 Feb 21. doi: 10.3791/2513
PMCID: PMC3197407
Isolation of Mouse Respiratory Epithelial Cells and Exposure to Experimental Cigarette Smoke at Air Liquid Interface
Hilaire C. Lam, Augustine M.K. Choi, and Stefan W. Ryter
Author information ► Copyright and License information ►
This article has been cited by other articles in PMC.
Go to:
Abstract
Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated respiratory epithelium. A protocol is described for isolating and exposing these cells to mainstream cigarette smoke (CS), in order to study epithelial cell responses to CS exposure. The protocol consists of three parts: the isolation of airway epithelial cells from mouse trachea, the culturing of these cells at air-liquid interface (ALI) as fully differentiated epithelial cells, and the delivery of calibrated mainstream CS to these cells in culture. The ALI culture system allows the culture of respiratory epithelia under conditions that more closely resemble their physiological setting than ordinary liquid culture systems. The study of molecular and lung cellular responses to CS exposure is a critical component of understanding the impact of environmental air pollution on human health. Research findings in this area may ultimately contribute towards understanding the etiology of chronic obstructive pulmonary disease (COPD), and other tobacco-related diseases, which represent major global health problems.h
Dear PB,
I hope you can figure out any surface CD and its respective antibody for epithelial cells. later either you can sort cells by FACS and then culture on normal plates.
I hope this would be of help.
Regards
Prasun
Thank u so much for ur suggetion bt, what I m following is much simpler protocol without using any ab
- Badges
- Science topic
- Similar topics
- Chemistry
- Pharmacology
- Medical Pharmacology
- Clinical Pharmacology