Ultrasonic-augmented Primary Adult Fibroblast Isolation
Author(s) -
Stephan R. Künzel,
Charlotte Schaeffer,
Karolina Sekeres,
Carola S. Mehnert,
Stephanie M. Schacht Wall,
Manja Newe,
Susanne Kämmerer,
Ali ElArmouche
Publication year - 2019
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/59858
Subject(s) - ultrasonic sensor , fibroblast , isolation (microbiology) , microbiology and biotechnology , biology , medicine , pathology , cell culture , bioinformatics , genetics , radiology
Primary adult fibroblasts have become an important tool to study fibrosis, fibroblast interactions and inflammation in all body tissues. Since primary fibroblasts cannot divide indefinitely due to myofibroblast differentiation or senescence induction, new cultures must be established regularly. However, there are several obstacles to overcome during the processes of developing a reliable isolation protocol and primary fibroblast isolation itself: the method's degree of difficulty (especially for beginners), the risk of bacterial contamination, the required time until primary fibroblasts can be used for experiments, and subsequent cell quality and viability. In this study, a fast, reliable and easy-to-learn protocol to isolate and culture primary adult fibroblasts from mouse heart, lung, liver and kidney combining enzymatic digestion and ultrasonic agitation is provided.
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