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A High Output Method to Isolate Cerebral Pericytes from Mouse
Published on: January 14, 2020
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A High Output Method to Isolate Cerebral Pericytes from Mouse.
Anupriya Mehra1, Lucie Dehouck1, Elodie Vandenhaute1
1Laboratory of the Blood Brain Barrier, University of Artois.
Journal of Visualized Experiments : Jove
|February 4, 2020
Summary
Researchers developed a new, efficient method for isolating cerebral pericytes, crucial cells for brain barrier function. This antibiotic-free protocol significantly increases pericyte yield, reducing animal use and advancing neurological disorder research.
Area of Science:
- Vascular Biology
- Neuroscience
- Cell Biology
Background:
- Cerebral pericytes are vital for blood-brain barrier (BBB) formation and function.
- Their precise molecular roles in BBB physiology and neurological disorders require further investigation.
- Current in vitro models for cerebral pericyte extraction often lack high output and antibiotic-free conditions.
Purpose of the Study:
- To present a simple, efficient, and high-yield method for extracting cerebral pericytes.
- To provide an antibiotic-free protocol that minimizes animal usage.
- To establish a reliable in vitro model for studying pericyte physiology and pathology.
Main Methods:
- Homogenization of mouse brain tissue.
- Separation of microvascular fragments using a BSA-dextran solution.
- A three-step separation process followed by filtration to obtain microvessel-rich filtrate.
- Confirmation of pericyte purity using markers NG2, PDGFR-β, and CD146.
Main Results:
- The method yields sufficient microvascular fragments from 10 mice to seed 9 wells of a 6-well plate.
- The protocol enables the obtainment of 27 pericyte-rich wells in passage 2.
- High purity of pericyte cultures was confirmed via specific marker expression.
Conclusions:
- This novel method offers an efficient and feasible in vitro tool for cerebral pericyte research.
- The high output and antibiotic-free nature of the protocol are advantageous for physiological and pathophysiological studies.
- This technique facilitates deeper understanding of pericyte roles in neurological disorders.
