Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies

Isolation of fresh endothelial cells from porcine heart for cardiovascular studies: a new fast protocol suitable for genomic, transcriptomic and cell biology studies

Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is commonly used in human cli...

Full description

Saved in:
Bibliographic Details
Published in:BMC molecular and cell biology Vol. 20; no. 1; p. 32
Main Authors: Hätinen, Olli-Pekka A, Lähteenvuo, Johanna E, Korpela, Henna J, Pajula, Juho J, Ylä-Herttuala, Seppo
Format: Journal Article
Language:English
Published: England BioMed Central Ltd 13-08-2019
BioMed Central
BMC
Subjects:
Analysis
Animals
Cardiovascular Diseases - pathology
Cell Biology
Cell Separation - methods
Cells, Cultured
Endothelial cell
Endothelial Cells - cytology
Endothelium
Female
Fresh tissue
Genomics
Identification and classification
Isolation
Methodology
Methods
Myocardial
Myocardium - cytology
Neovascularization
Pig
Porcine heart
Separation (Technology)
Swine
Transcriptome - genetics
Finland
Isolation
Endothelial cell
Fresh tissue
Porcine heart
Myocardial
Pig
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Endothelial cells (ECs) play a key role in tissue homeostasis, in several pathological conditions, and specifically in the control of vascular functions. ECs are frequently used as in vitro model systems for cardiovascular studies and vascular biology. The porcine model is commonly used in human clinical cardiovascular studies. Currently, however, there is no robust protocol for the isolation of porcine heart ECs. We have developed a fast isolation protocol, which is cost effective, takes only 1-2 h, and produces EC purity of over 97%. This protocol is optimized for porcine hearts but can be adapted for use with other large animals. Heart is washed by flushing with PBS, whereafter endothelial cells are detached by collagenase incubation and the cells can then be collected immediately after the incubation and plated within an hour after the heart is isolated from a pig. The swiftness of the protocol limits changes in the phenotype and RNA expression profile of the cells. Cells were identified as ECs with CD31 (PECAM-1) antibody immunostaining. Functionality of ECs were ensured with in vitro angiogenesis assay. The purity of the ECs was verified by using fluorescence assisted cell sorting (FACS) with the CD31 antibody. We developed a new, fast, and cost-effective isolation method for pig heart ECs. Successful isolation of pure ECs is a prerequisite for several cardiovascular and vascular biology studies.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:2661-8850
2661-8850
DOI:10.1186/s12860-019-0215-2