[Isolation, culture and identification of satellite cells in rat multifidus muscle]

Objective To establish an efficient protocol for isolation, culture and identification of satellite cells in rat multifidus muscle. Methods After deep anesthesia, lumber 4 and 5 multifidus muscle were isolated from 4-week postnatal female SD rats and digested with collagenase 1. Then, the satellite cells were purified via differential adhesion velocity. They were characterized by the detection of marker proteins such as paired box 7 (Pax7), myogenic differentiation (MyoD) and myosin heavy chain (MyHC) with immunofluorescence staining. Results After isolation and purification, satellite cells were found to be circular in morphology and with high optical refraction. They became spindle-shaped after cultured for 5 days. The isolated satellite cells had similar growth style and morphological characteristics. They expressed Pax7 at the interphase of mitosis, MyoD at the proliferating phase and MyHC at the early differentiation stage. Conclusion This study successfully established a simple and efficient method of isolation, culture and identification of satellite cells in rat multifidus muscle.