Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line

Mohd Nor, Nor Hafizah and Mohamed, Farahidah and Mohd Shafri, Mohd Affendi (2014) Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line. In: K.A.H.S. Research Week 2014, 17-21 Nov. 2014, KAHS IIUM, Kuantan, Pahang.

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Abstract

INTRODUCTION: The BBB constitutes the major obstacle to drug delivery to the brain. To test BBB’s permeability using the paracellular, non-specific transport pathway of a newlyformulated hydrophilic particle for neuromodulating application, BBB model using the EOO (epithelial cell/absent cell/absent cell) arrangement was constructed. MATERIALS AND METHODS: Briefly, Madin-Darby Canine Kidney (MDCK) 1 cells were cultured in Dulbecco’s Minimum Essential Medium (DMEM) complete growth media and plated at the density of 1 x 105 on 0.4 μm PET filter of the Millipore hanging cell culture inserts supplemented with 0.3 ml of the complete growth medium in the luminal space and 1.1 ml of the complete growth media in the abluminal space. The inserts were placed in the 24-well plates to replicate the parenchymal and microvascular space of the BBB. Media were changed for every 2 alternate days, cells were allowed to resume growth and differentiate within 4-9 days, and finally subjected to the permeation study involving the ‘sink condition’ where the complete growth media were substituted with Hanks Balance Salt Solution (HBSS) pH 7.4 (0.2 ml in the luminal space; 1.1 ml in the abluminal space) for pre-equilibration for 1 hour. Cells were moderately shaken in the incubator shaker for 30 minutes at 100rpm, and trans-epithelial electrical resistance (TEER) reading was measured every 30 minutes up to 3 hours. Then, cells were washed with HBSS and substituted with the complete culture media before being stored overnight in the incubator (37ºC, 95% relative humidity, 5% CO2). The recovery of the TEER was measured 24 hours post-treatment using similar measurement used at the start of the experiment. RESULTS AND DISCUSSION: In this study, the TEER values of the MDCK cells were in the range of ~600 to ~900 Ω x cm2 . CONCLUSION: Hence the integrity of MDCK 1 BBB was successfully sustained to meet standard models.

Item Type:	Conference or Workshop Item (Poster)
Additional Information:	4341/39613
Uncontrolled Keywords:	Blood-brain barrier (BBB), neuromodulation, MDCK-1 cell line, permeation test
Subjects:	R Medicine > R Medicine (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Allied Health Sciences > Department of Biomedical Science (Effective:1st July 2011)
Depositing User:	Dr Mohd Affendi Mohd Shafri
Date Deposited:	12 Jan 2015 11:30
Last Modified:	06 Jul 2015 08:55
URI:	http://irep.iium.edu.my/id/eprint/39613

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