Adherent Primary Cultures of Mouse Intercostal Muscle Fibers for Isolated Fiber Studies
Figure 2
Dissociation. Following digestion, ribs are transferred into MEM containing 10% FBS and gentamycin (a). Rib pair flanking the desired muscle (white arrows) is isolated (b) and transferred into clean 60 mm Falcon Petri dish containing 7 mL MEM with 10% FBS and gentamycin. Fibers are triturated (c) until a significant population of fibers are isolated (d). Fibers may be aggregated in the center of dish (e) by shaking gently in a circular motion. FinnTip 250 Wide pipette tips are used to plate fibers (f) to prevent damage from fluidic shearing. Brightfield images of an adhered fiber at low (g) and high (h) magnification. Scale bars are approximately 1 mm (d, e) or 50 μm (g, h).
