Isolation and Transfection of Primary Culture Bovine Retinal Pericytes

Vincent A Primo; Joseph F Arboleda-Velasquez

doi:10.1007/978-1-4939-3628-1_7

Isolation and Transfection of Primary Culture Bovine Retinal Pericytes

Methods Mol Biol. 2016:1430:107-17. doi: 10.1007/978-1-4939-3628-1_7.

Authors

Vincent A Primo¹, Joseph F Arboleda-Velasquez²

Affiliations

¹ Department of Ophthalmology, Schepens Eye Research Institute, Mass Eye and Ear Infirmary, Harvard Medical School, 20 Staniford Street, Boston, MA, 02114, USA.
² Department of Ophthalmology, Schepens Eye Research Institute, Mass Eye and Ear Infirmary, Harvard Medical School, 20 Staniford Street, Boston, MA, 02114, USA. Joseph_Arboleda@meei.harvard.edu.

PMID: 27172949
DOI: 10.1007/978-1-4939-3628-1_7

Abstract

This protocol describes an enzymatic approach for isolating homogeneous cultures of pericytes from retinas of bovine source. In summary, retinas are dissected, washed, digested, filtered, cultured in specific media to select for pericytes, and finally expanded for a low passage culture of about 14 million bovine retinal pericytes (BRP) within 4-6 weeks. This protocol also describes a liposomal-based technique for transfection of BRPs.

Keywords: Bovine; Isolation; Pericyte; Primary culture; Retina.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cattle
Cell Culture Techniques
Cell Proliferation
Cell Separation / methods*
Cells, Cultured
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism
Liposomes / metabolism
Pericytes / cytology*
Plasmids / genetics
Retinal Vessels / cytology*
Transfection / methods*

Substances

Liposomes
Green Fluorescent Proteins

Grants and funding

K99 EY021624/EY/NEI NIH HHS/United States