Biological characteristics of muscle-derived satellite cells isolated from rats at different postnatal days

Abstract

This study investigated the in vitro growth characteristics and differential potential of musclederived satellite cells (MDSCs) derived from rats at different postnatal (P) stages, in order to expand the range of source material for tissue engineering. Rat MDSCs were isolated from P5, P10, P15, P21 and P42 rat skeletal muscles using double enzyme digestion and differential adherent culture. Neurogenic, osteogenic and myogenic induction media were used to induce directed differentiation. Differentiated nerve cells, osteoblasts and myotubes were identified by their morphology and immunohistochemical staining. Most cells transformed into spindle-shaped mononuclear cells after 48 h and proliferated rapidly. MDSCs were difficult to isolate from P42 rats. After neurogenesis, four groups MDSCs formed neuron-specific enolase positive polygonal-shaped dendritic cells. After osteogenesis, P5, P10, P15 and P21 MDSCs formed Alizarin red-and osteocalcin-positive bone nodules. After myogenesis, myotubes were formed and were fast muscle myosin-positive. MDSCs derived from P5, P10, P15 and P21 rat skeletal muscle are easy to isolate, culture and amplify in vitro, which increases the range of source material available for tissue engineering.

Differentially expressed microRNAs during the differentiation of muscle-derived stem cells into insulin-producing cells, a promoting role of microRNA-708-5p/STK4 axis

“…MDSCs were isolated from the leg muscle of Wistar rats according to the previously described method [14]. Briefly, the muscle tissues were collected from Wistar rats and digested with 0.1% type I collagenase for 1 h at 37˚C.…”

Withdrawal: Muscle‐derived Satellite Cells for Treating Type 1 Diabetes in Rats (Rattus norvegicus)

“…Rat MDSCs were isolated using the method we have previously described (Yu et al, 2015). Briefly, after disinfection, the muscles were cut into 1-2 mm 2 fragments, digested with 0.1% type I collagenase in phosphate-buffered saline (PBS) for 1 hr at 37°C in a water bath with shaking, and centrifuged at 150 g for 5 min.…”

“…The cells were plated in culture dishes coated with 0.1% gelatin at a density of 1 × 10 6 cells/ml. MDSCs were purified using the differential adhesion method (Ren et al, 2014;Yu et al, 2015), and cultured in growth medium. Eighty percent confluent 3rd generation MDSCs were cultured in myogenic induction medium (2% HS + 98% DMEM/F12) for 5 days.…”