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

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/abs...

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Main Authors:	Mohd Nor, Nor Hafizah, Mohamed, Farahidah, Mohd Shafri, Mohd Affendi
Format:	Conference or Workshop Item
Language:	English English
Published:	2014
Subjects:	R Medicine (General)
Online Access:	http://irep.iium.edu.my/39613/ http://irep.iium.edu.my/39613/1/Poster_17.pdf http://irep.iium.edu.my/39613/4/39613.pdf

id	iium-39613
recordtype	eprints
spelling	iium-396132015-07-06T00:55:55Z http://irep.iium.edu.my/39613/ Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line Mohd Nor, Nor Hafizah Mohamed, Farahidah Mohd Shafri, Mohd Affendi R Medicine (General) 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. 2014 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/39613/1/Poster_17.pdf application/pdf en http://irep.iium.edu.my/39613/4/39613.pdf 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.
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	R Medicine (General)
spellingShingle	R Medicine (General) Mohd Nor, Nor Hafizah Mohamed, Farahidah Mohd Shafri, Mohd Affendi Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
description	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.
format	Conference or Workshop Item
author	Mohd Nor, Nor Hafizah Mohamed, Farahidah Mohd Shafri, Mohd Affendi
author_facet	Mohd Nor, Nor Hafizah Mohamed, Farahidah Mohd Shafri, Mohd Affendi
author_sort	Mohd Nor, Nor Hafizah
title	Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
title_short	Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
title_full	Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
title_fullStr	Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
title_full_unstemmed	Construction of in vitro Blood-Brain Barrier (BBB) model using MDCK-1 cell line
title_sort	construction of in vitro blood-brain barrier (bbb) model using mdck-1 cell line
publishDate	2014
url	http://irep.iium.edu.my/39613/ http://irep.iium.edu.my/39613/1/Poster_17.pdf http://irep.iium.edu.my/39613/4/39613.pdf
first_indexed	2023-09-18T20:56:56Z
last_indexed	2023-09-18T20:56:56Z
_version_	1777410366658052096

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

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