Choroid Plexus Epithelial Monolayers--a Cell Culture Model from Porcine Brain

Overview

Journal Cerebrospinal Fluid Res

Publisher Biomed Central

Date 2006 Dec 23

PMID 17184532

Citations 28

Authors

Carsten Baehr

Valeska Reichel

Gert Fricker

Affiliations

Soon will be listed here.

Abstract

Background: The goal of the present study was to develop an in vitro choroid plexus (CP) epithelial cell culture model for studying transport of protein-mediated drug secretion from blood to cerebrospinal fluid (CSF) and vice versa.

Methods: Cells were isolated by mechanical and enzymatic treatment of freshly isolated porcine plexus tissue. Epithelial cell monolayers were grown and CSF secretion and transepithelial resistance were determined. The expression of f-actin as well as the choroid plexus marker protein transthyretin (TTR), were assessed. The expression of the export proteins p-glycoprotein (Pgp, Abcb1) and multidrug resistance protein 1 (Mrp1, Abcc1) was studied by RT-PCR, Western-blot and immunofluorescence techniques and their functional activity was assessed by transport and uptake experiments.

Results: Choroid plexus epithelial cells were isolated in high purity and grown to form confluent monolayers. Filter-grown monolayers displayed transendothelial resistance (TEER) values in the range of 100 to 150 ohms cm2. Morphologically, the cells showed the typical net work of f-actin and expressed TTR at a high rate. The cultured cells were able to secrete CSF at a rate of 48.2 +/- 4.6 microl/cm2/h over 2-3 hours. The ABC-export protein Mrp1 was expressed in the basolateral (blood-facing) membranes of cell monolayers and intact tissue. P-glycoprotein showed only low expression within the apical (CSF directed) membrane but was located more in sub-apical cell compartments. This finding was paralleled by the lack of directed excretion of p-glycoprotein substrates, verapamil and rhodamine 123.

Conclusion: It was demonstrated that CP epithelium can be isolated and cultured, with cells growing into intact monolayers, fully differentiating and with properties resembling the tissue in vivo. Thus, the established primary porcine CP model, allowing investigation of complex transport processes, can be used as a reliable tool for analysis of xenobiotic transport across the blood-cerebrospinal fluid barrier (BCSFB).

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References

Strazielle N, Ghersi-Egea J . Demonstration of a coupled metabolism-efflux process at the choroid plexus as a mechanism of brain protection toward xenobiotics. J Neurosci. 1999; 19(15):6275-89. PMC: 6782833. View

Breen C, Sykes D, Baehr C, Fricker G, Miller D . Fluorescein-methotrexate transport in rat choroid plexus analyzed using confocal microscopy. Am J Physiol Renal Physiol. 2004; 287(3):F562-9. DOI: 10.1152/ajprenal.00045.2003. View

Wegener J, Sieber M, Galla H . Impedance analysis of epithelial and endothelial cell monolayers cultured on gold surfaces. J Biochem Biophys Methods. 1996; 32(3):151-70. DOI: 10.1016/0165-022x(96)00005-x. View

Anderson J, Van Itallie C . Tight junctions and the molecular basis for regulation of paracellular permeability. Am J Physiol. 1995; 269(4 Pt 1):G467-75. DOI: 10.1152/ajpgi.1995.269.4.G467. View

Hakvoort A, Haselbach M, Galla H . Active transport properties of porcine choroid plexus cells in culture. Brain Res. 1998; 795(1-2):247-56. DOI: 10.1016/s0006-8993(98)00284-4. View

Lee G, Dallas S, Hong M, Bendayan R . Drug transporters in the central nervous system: brain barriers and brain parenchyma considerations. Pharmacol Rev. 2001; 53(4):569-96. View

Ghersi-Egea J, Strazielle N . Brain drug delivery, drug metabolism, and multidrug resistance at the choroid plexus. Microsc Res Tech. 2001; 52(1):83-8. DOI: 10.1002/1097-0029(20010101)52:1<83::AID-JEMT10>3.0.CO;2-N. View

Hilden S, Johns C, Guggino W, Madias N . Techniques for isolation of brush-border and basolateral membrane vesicles from dog kidney cortex. Biochim Biophys Acta. 1989; 983(1):77-81. DOI: 10.1016/0005-2736(89)90382-9. View

Mayer S, Sanders-Bush E . Sodium-dependent antiporters in choroid plexus epithelial cultures from rabbit. J Neurochem. 1993; 60(4):1308-16. DOI: 10.1111/j.1471-4159.1993.tb03291.x. View

10.

Fricker G, Miller D . Modulation of drug transporters at the blood-brain barrier. Pharmacology. 2004; 70(4):169-76. DOI: 10.1159/000075545. View

11.

Begley D . ABC transporters and the blood-brain barrier. Curr Pharm Des. 2004; 10(12):1295-312. DOI: 10.2174/1381612043384844. View

12.

Saito Y, Wright E . Bicarbonate transport across the frog choroid plexus and its control by cyclic nucleotides. J Physiol. 1983; 336:635-48. PMC: 1198989. DOI: 10.1113/jphysiol.1983.sp014602. View

13.

Stauder A, Dickson P, Aldred A, Schreiber G, Mendelsohn F, Hudson P . Synthesis of transthyretin (pre-albumin) mRNA in choroid plexus epithelial cells, localized by in situ hybridization in rat brain. J Histochem Cytochem. 1986; 34(7):949-52. DOI: 10.1177/34.7.3458812. View

14.

Redzic Z, Isakovic A, Misirlic Dencic S, Popadic D, Segal M . Uneven distribution of nucleoside transporters and intracellular enzymatic degradation prevent transport of intact [14C] adenosine across the sheep choroid plexus epithelium as a monolayer in primary culture. Cerebrospinal Fluid Res. 2006; 3:4. PMC: 1450313. DOI: 10.1186/1743-8454-3-4. View

15.

Miller D, Nobmann S, Gutmann H, Toeroek M, Drewe J, Fricker G . Xenobiotic transport across isolated brain microvessels studied by confocal microscopy. Mol Pharmacol. 2000; 58(6):1357-67. DOI: 10.1124/mol.58.6.1357. View

16.

Rao V, Dahlheimer J, Bardgett M, Snyder A, Finch R, Sartorelli A . Choroid plexus epithelial expression of MDR1 P glycoprotein and multidrug resistance-associated protein contribute to the blood-cerebrospinal-fluid drug-permeability barrier. Proc Natl Acad Sci U S A. 1999; 96(7):3900-5. PMC: 22392. DOI: 10.1073/pnas.96.7.3900. View

17.

Gath U, Hakvoort A, Wegener J, Decker S, Galla H . Porcine choroid plexus cells in culture: expression of polarized phenotype, maintenance of barrier properties and apical secretion of CSF-components. Eur J Cell Biol. 1997; 74(1):68-78. View

18.

Crook R, Kasagami H, Prusiner S . Culture and characterization of epithelial cells from bovine choroid plexus. J Neurochem. 1981; 37(4):845-54. DOI: 10.1111/j.1471-4159.1981.tb04470.x. View

19.

Scheffer G, Kool M, Heijn M, de Haas M, Pijnenborg A, Wijnholds J . Specific detection of multidrug resistance proteins MRP1, MRP2, MRP3, MRP5, and MDR3 P-glycoprotein with a panel of monoclonal antibodies. Cancer Res. 2000; 60(18):5269-77. View

20.

De Bault L, Mitro A . Species differences in the distribution of gamma-glutamyl transpeptidase in choroid plexus of lateral ventricle and microvessels of adjacent brain. Histochem J. 1994; 26(5):447-52. DOI: 10.1007/BF00160058. View