Abstract
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.
Highlights
Pipetus SafeSeal tips for pipettes (10 μl, 20 μl, 100 μl, 200 μl, 1000 μl) 5 ml, 10 ml, 25 ml, 50 ml Techno cut High glucose
MS2 Minishaker (subsequent model: Ident-Nr.: 0020016017) BD Falcon tubes (15 ml, 50 ml)
Summary
Pipetus SafeSeal tips for pipettes (10 μl, 20 μl, 100 μl, 200 μl, 1000 μl) 5 ml, 10 ml, 25 ml, 50 ml Techno cut High glucose. Materials List for: Ultrasonic-Augmented Primary Adult Fibroblast Isolation Kuenzel1, Charlotte Schaeffer1, Karolina Sekeres1, Carola S.
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