Isolation and characterization of peritoneal microvascular pericytes
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Updated: Oct 2, 2025
Isolation and Purification of Murine Cardiac Pericytes
Published on: August 16, 2019
Lei Tang1,2, Jun Shi3, Manshu Yu2
1First Clinic Medical School, Nanjing University of Chinese Medicine, China.
Abstract:
As a potential source of myofibroblasts, pericytes may play a role in human peritoneal fibrosis. The culture of primary vascular pericytes in animals has previously been reported, most of which are derived from cerebral and retinal microvasculature. Here, in the field of peritoneal dialysis, we describe a method to isolate and characterize mouse peritoneal microvascular pericytes. The mesenteric tissues of five mice were collected and digested by type II collagenase and type I DNase. After cell attachment, the culture fluid was replaced with pericyte-conditioned medium. Pericytes with high purity (99.0%) could be isolated by enzymatic disaggregation combined with conditional culture and magnetic activated cell sorting. The primary cells were triangular or polygonal with protrusions, and confluent cell culture could be established in 3 days. The primary pericytes were positive for platelet-derived growth factor receptor-β, α-smooth muscle actin, neuron-glial antigen 2, and CD13. Moreover, they promoted formation of endothelial tubes, and pericyte-myofibroblast transition occurred after treatment with transforming growth factor-β1. In summary, we describe here a reproducible isolation protocol for primary peritoneal pericytes, which may be a powerful tool for in vitro peritoneal fibrosis studies.
Insights
Researchers developed a method to isolate mouse peritoneal pericytes, crucial cells for studying peritoneal fibrosis in dialysis patients. This technique yields highly pure pericytes for in vitro research.
Area of Science:
- Cell Biology
- Biomedical Engineering
- Nephrology
Background:
- Pericytes are implicated as a source of myofibroblasts in human peritoneal fibrosis.
- Previous studies on primary vascular pericyte culture primarily focused on cerebral and retinal microvasculature.
Purpose of the Study:
- To establish a reproducible method for isolating and characterizing mouse peritoneal microvascular pericytes.
- To provide a tool for in vitro studies of peritoneal fibrosis.
Main Methods:
- Isolation of mouse peritoneal microvascular pericytes from mesenteric tissues using enzymatic digestion (collagenase II, DNase I).
- Culturing cells in pericyte-conditioned medium after initial attachment.
- Purification using magnetic activated cell sorting (MACS).
Main Results:
- Achieved high purity (99.0%) of primary peritoneal pericytes.
- Characterized cells morphologically (triangular/polygonal) and confirmed purity via positive staining for PDGFR-β, α-SMA, NG2, and CD13.
- Demonstrated pericyte function in promoting endothelial tube formation and pericyte-myofibroblast transition upon TGF-β1 stimulation.
Conclusions:
- A reproducible protocol for isolating primary mouse peritoneal pericytes was successfully developed.
- These isolated pericytes serve as a valuable tool for in vitro research into peritoneal fibrosis mechanisms.
- The findings contribute to understanding the role of pericytes in peritoneal fibrosis, particularly in the context of peritoneal dialysis.
