Isolation and culture of endothelial cells, pericytes and perivascular resident macrophage-like melanocytes from the young mouse ear

Nature Protocols

Volumn 8, Issue 4, 2013, Pages 709-720

  Neng, Lingling ^a,b   Zhang, Wenjing ^a,b   Hassan, Ahmed ^c   Zemla, Marcin ^c   Kachelmeier, Allan ^a   Fridberger, Anders ^d   Auer, Manfred ^c   Shi, Xiaorui ^a  

^a OREGON HEALTH AND SCIENCE UNIVERSITY   (United States)

^b THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIVERSITY   (China)

^c LAWRENCE BERKELEY NATIONAL LABORATORY   (United States)

^d KAROLINSKA INSTITUTE   (Sweden)

Author keywords

[No Author keywords available]

Indexed keywords

PIGMENT EPITHELIUM DERIVED FACTOR;

ANIMAL TISSUE; ARTICLE; CELL CULTURE; CELL INTERACTION; CELL ISOLATION; CULTURE MEDIUM; CULTURE TECHNIQUE; ENDOTHELIUM CELL; EXPLANT; INTERMETHOD COMPARISON; MELANOCYTE; MOUSE; NONHUMAN; PERICYTE; PERIVASCULAR RESIDENT MACROPHAGE LIKE MELANOCYTE; PRIORITY JOURNAL; STRIA VASCULARIS;

ANIMALS; CELL CULTURE TECHNIQUES; CELL LINE; CELL SEPARATION; CRYOPRESERVATION; CULTURE MEDIA; EAR; ENDOTHELIAL CELLS; FEMALE; MACROPHAGES; MALE; MELANOCYTES; MICE; MICE, INBRED C57BL; PERICYTES; STRIA VASCULARIS;

MUS;

EID: 84875496365     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2013.033     Document Type: Article

References (33)

1. Cohen-Salmon, M. et al. Connexin30 deficiency causes instrastrial fluid-blood barrier disruption within the cochlear stria vascularis. Proc. Natl. Acad. Sci. USA 104, 6229-6234 (2007).
2. Juhn, S.K., Hunter, B.A. & Odland, R.M. Blood-labyrinth barrier and fluid dynamics of the inner ear. Int. Tinnitus J. 7, 72-83 (2001).
3. Juhn, S.K. & Rybak, L.P. Labyrinthine barriers and cochlear homeostasis. Acta Otolaryngol. 91, 529-534 (1981).
4. Juhn, S.K., Rybak, L.P. & Fowlks, W.L. Transport characteristics of the blood-perilymph barrier. Am. J. Otolaryngol. 3, 392-396 (1982).
5. Zhang, W. et al. Perivascular-resident macrophage-like melanocytes in the inner ear are essential for the integrity of the intrastrial fluid-blood barrier. Proc. Natl. Acad. Sci. USA 109, 10388-10393 (2012).
6. Cucullo, L. et al. A new dynamic in vitro model for the multidimensional study of astrocyte-endothelial cell interactions at the blood-brain barrier. Brain Res. 951, 243-254 (2002).
7. Duport, S. et al. An in vitro blood-brain barrier model: cocultures between endothelial cells and organotypic brain slice cultures. Proc. Natl. Acad. Sci. USA 95, 1840-1845 (1998).
8. Lai, C.H. & Kuo, K.H. The critical component to establish in vitro BBB model: pericyte. Brain Res. Brain Res. Rev. 50, 258-265 (2005).
9. Folkman, J., Haudenschild, C.C. & Zetter, B.R. Long-term culture of capillary endothelial cells. Proc. Natl. Acad. Sci. USA 76, 5217-5221 (1979).
10. Baudin, B., Bruneel, A., Bosselut, N. & Vaubourdolle, M. A protocol for isolation and culture of human umbilical vein endothelial cells. Nat. Protoc. 2, 481-485 (2007).
11. Jaffe, E.A., Nachman, R.L., Becker, C.G. & Minick, C.R. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J. Clin. Invest. 52, 2745-2756 (1973).
12. Sobczak, M., Dargatz, J. & Chrzanowska-Wodnicka, M. Isolation and culture of pulmonary endothelial cells from neonatal mice. J. Vis. Exp. 46, 2316 (2010).
13. Bernas, M.J. et al. Establishment of primary cultures of human brain microvascular endothelial cells to provide an in vitro cellular model of the blood-brain barrier. Nat. Protoc. 5, 1265-1272 (2010).
14. Maier, C.L., Shepherd, B.R., Yi, T. & Pober, J.S. Explant outgrowth, propagation and characterization of human pericytes. Microcirculation 17, 367-380 (2010).
15. Mogensen, C. et al. Isolation and functional characterization of pericytes derived from hamster skeletal muscle. Acta Physiol. 201, 413-426 (2011).
16. Weber, S.C. et al. Isolation and culture of fibroblasts, vascular smooth muscle, and endothelial cells from the fetal rat ductus arteriosus. Pediatr. Res. 70, 236-241 (2011).
17. Bobilya, D.J. A model for transport studies of the blood-brain barrier. Methods Mol. Biol. 637, 149-163 (2010).
18. Bryan, B.A. & D′Amore, P.A. Pericyte isolation and use in endothelial/pericyte coculture models. Methods Enzymol. 443, 315-331 (2008).
19. Rops, A.L. et al. Isolation and characterization of conditionally immortalized mouse glomerular endothelial cell lines. Kidney Int. 66, 2193-2201 (2004).
20. Ribatti, D., Nico, B., Vacca, A., Roncali, L. & Dammacco, F. Endothelial cell heterogeneity and organ specificity. J. Hematother. Stem Cell Res. 11, 81-90 (2002).
21. Seaman, S. et al. Genes that distinguish physiological and pathological angiogenesis. Cancer cell 11, 539-554 (2007).
22. Neng, L., Zhang, F., Kachelmeier, A. & Shi, X. Endothelial cell, pericyte, and perivascular resident macrophage-type melanocyte interactions regulate cochlear intrastrial fluid-blood barrier permeability. J. Assoc. Res. Otolaryngol. http://dx.doi.org/10.1007/s10162-012-0365-9 (2012).
23. Iwagaki, T., Suzuki, T. & Nakashima, T. Development and regression of cochlear blood vessels in fetal and newborn mice. Hear. Res. 145, 75-81 (2000).
24. Shi, X. Resident macrophages in the cochlear blood-labyrinth barrier and their renewal via migration of bone-marrow-derived cells. Cell Tissue Res. 342, 21-30 (2010).
25. Tigges, U., Welser-Alves, J.V., Boroujerdi, A. & Milner, R. A novel and simple method for culturing pericytes from mouse brain. Microvasc. Res. 84, 74-80 (2012).
26. Wisniewska-Kruk, J. et al. A novel co-culture model of the blood-retinal barrier based on primary retinal endothelial cells, pericytes and astrocytes. Exp. Eye Res. 96, 181-190 (2012).
27. Yamagishi, S. et al. Pigment epithelium-derived factor protects cultured retinal pericytes from advanced glycation end product-induced injury through its antioxidative properties. Biochem. Biophys. Res. Commun. 296, 877-882 (2002).
28. Dawson, D.W. et al. Pigment epithelium-derived factor: a potent inhibitor of angiogenesis. Science 285, 245-248 (1999).
29. Shepro, D. & Morel, N.M. Pericyte physiology. FASEB J. 7, 1031-1038 (1993).
30. Vandenhaute, E. et al. Brain pericytes from stress-susceptible pigs increase blood-brain barrier permeability. Fluids Barriers CNS 9, 11 (2012).
31. Bouchard, B.A., Shatos, M.A. & Tracy, P.B. Human brain pericytes differentially regulate expression of procoagulant enzyme complexes comprising the extrinsic pathway of blood coagulation. Arterioscler. Thromb. Vasc. Biol. 17, 1-9 (1997).
32. Provance, D.W. Jr., Wei, M., Ipe, V. & Mercer, J.A. Cultured melanocytes from dilute mutant mice exhibit dendritic morphology and altered melanosome distribution. Proc. Natl. Acad. Sci. USA 93, 14554-14558 (1996).
33. Gerrity, R.G., Richardson, M., Somer, J.B., Bell, F.P. & Schwartz, C.J. Endothelial cell morphology in areas of in vivo Evans blue uptake in the aorta of young pigs. II. Ultrastructure of the intima in areas of differing permeability to proteins. Am. J. Pathol. 89, 313-334 (1977).