Related Experiment Video
Updated: May 3, 2026
Isolation and Culture of Primary Retinal Müller Cells from Sprague-Dawley (SD) Rats
Published on: June 17, 2025
Isolation, purification, and cultivation of primary retinal microvascular pericytes: a novel model using rats
Guanghui Liu1, Chun Meng, Mingdong Pan
1School of Optometry and Ophthalmology and Eye Hospital, Wenzhou Medical University, Wenzhou, China; Department of Ophthalmology, Affiliated People's Hospital (People's Hospital of Fujian Province), Fujian University of Traditional Chinese Medicine, Fuzhou, China.
Objective:
To isolate, purify, and cultivate primary retinal microvascular pericytes (RMPs) from rats to facilitate the study of their properties in vitro.
Methods:
Primary RMPs were isolated from weanling rats by mechanical morcel and collagenase digestion, and purified by a step-wise combination of selective medium with different glucose concentrations, medium exchange, and partial enzymatic digestion. Morphology of RMPs was assessed by phase contrast microscopy. Further characterization was analyzed by immunofluorescence. Functional assay was evaluated by the pericytes- endothelial cells (ECs) coculture system.
Results:
Retinal microvascular pericytes migrated out of microvascular fragments after 24-48 hours of plating and reached subconfluence on days 14-16. The cells showed typical pericyte morphology with large irregular triangular cell bodies and multiple long processes, and uniformly expressed the cellular markers α-SMA, PDGFR-β, NG2 and desmin, but were negative for vWF, GS, GFAP and SMMHC. Ninety-nine percent of the cell population had double positive staining for α-SMA and PDGFR-β. In the coculture system, RMPs can directly contact ECs and move together to form the capillary-like cords.
Conclusions:
Retinal microvascular pericytes can be readily obtained by our method. We report the first cultivation of primary RMPs from rats and establish a simple method for their isolation and purification.
Insights
Researchers successfully isolated and cultivated primary retinal microvascular pericytes (RMPs) from rats. This new method enables further in vitro study of RMP properties and their role in retinal vasculature.
Area of Science:
- Ophthalmology
- Cell Biology
- Vascular Biology
Background:
- Retinal microvascular pericytes (RMPs) are crucial for maintaining blood-retinal barrier integrity and regulating retinal blood flow.
- Understanding RMP properties in vitro is essential for studying retinal vascular diseases.
Purpose of the Study:
- To develop a reliable method for isolating, purifying, and cultivating primary rat RMPs.
- To establish a foundation for in vitro studies on RMP function and behavior.
Main Methods:
- Primary RMPs were isolated from weanling rats using mechanical dissociation and collagenase digestion.
- Purification involved selective media, medium exchange, and partial enzymatic digestion.
- Cell characterization utilized phase contrast microscopy, immunofluorescence, and a co-culture system with endothelial cells (ECs).
Main Results:
- Isolated RMPs exhibited characteristic morphology and expressed key pericyte markers (α-SMA, PDGFR-β, NG2, desmin).
- Over 99% of cells were positive for α-SMA and PDGFR-β.
- In co-culture, RMPs interacted with ECs to form capillary-like structures.
Conclusions:
- A straightforward and effective method for obtaining primary rat RMPs was established.
- This study reports the first successful cultivation of primary rat RMPs, facilitating future research.
