Journal article
Purification of murine endothelial cell cultures by flow cytometry using fluorescein-labeled griffonia simplicifolia agglutinin
The American journal of pathology, Vol.134(6), pp.1227-1232
06/1989
PMCID: PMC1879935
PMID: 2757116
Abstract
Griffonia simplicifolia agglutinin (GSA) is a valuable histochemical tool in the identification of endothelium. In this study GSA labeled with fluorescein isothiocyanate (GSA-FITC) was used to purify cultures of murine cerebral microvascular endothelium. Cultures were stained with GSA-FITC, then sorted using a fluorescence-activated cell sorter (FACS). GSA-positive endothelial cells were collected, re-cultured, and subsequently re-analyzed by FACS using GSA-FITC. Cultures that initially contained 80 +/- 3 to 89 +/- 3% (X +/- SE) GSA-positive cells were purified to 98 +/- 1% positivity. Immunohistochemistry with an anti-muscle-action antibody confirmed that FACS sorting of GSA-FITC-stained cells effectively removed contaminating smooth muscle cells from endothelial cell cultures. Viability, proliferation, and prostaglandin production of the cells was unaltered by lectin staining and FACS sorting. Thus, GSA-FITC can be used in conjunction with flow cytometry to enhance the purity of murine endothelial cell cultures without adversely affecting cell viability, growth, or metabolism.
Details
- Title: Subtitle
- Purification of murine endothelial cell cultures by flow cytometry using fluorescein-labeled griffonia simplicifolia agglutinin
- Creators
- Geronimo Sahagun - Department of Pathology, University of Iowa, Iowa City 52242Steven A Moore - Department of Pathology, University of Iowa, Iowa City 52242Zsuzsanna Fabry - Department of Pathology, University of Iowa, Iowa City 52242Robert L Schelper - Department of Pathology, University of Iowa, Iowa City 52242Michael N Hart - Department of Pathology, University of Iowa, Iowa City 52242
- Resource Type
- Journal article
- Publication Details
- The American journal of pathology, Vol.134(6), pp.1227-1232
- PMID
- 2757116
- PMCID
- PMC1879935
- ISSN
- 0002-9440
- eISSN
- 1525-2191
- Language
- English
- Date published
- 06/1989
- Academic Unit
- Pathology
- Record Identifier
- 9984047632002771
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