I'm attempting to develop a BBB model in Transwell inserts using cryopreserved primary rat BMVECs; however, no significant TEER has developed. Most publications I've read use freshly isolated BMVECs to develop their BBB model, but I purchased frozen cells from Cell Applications through Sigma. Does freezing cells limit their ability to develop TEER and tight junctions? Has anyone seen a publication that uses frozen endothelial cells for a BBB model that successfully develop TEER?
Maybe Fenglin Shi I, and Kenneth L. Audus, Biochemical Characteristics of Primary and Passaged Cultures of Primate Brain Microvessel Endothelial Cells, Neurochem Res, Vol. 19, No. 4, 1994, pp. 427--433 could help. We always froze the isolated bovine brain capillary fragments after an isolation and seeded rat tail collagen-coated Transwells for expts with fairly consistent results. I suggest you reach out to Dr. Audus directly at [email protected]. He is the perfecter of the BMEC culture and a really nice guy as well. Say that I referred you. Good luck!
Frozen and thawed cells can develop sufficient electrical resistance.,
I recommend the follwoing article:
Hydrocortisone reinforces the blood-brain properties in a serum free cell culture system.Hoheisel et al. Biochem Biophys Res Commun. 1998 Jun 18;247(2):312-5.
Thank you, Gert Fricker ! I will take a look.
Thomas J Raub I appreciate your recommendation and will reach out to Dr. Audus. Thank you!
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