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Updated: Aug 25, 2026
Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells
Published on: September 21, 2010
Isolation of muscle derived stem cells from rat and its smooth muscle differentiation [corrected]
Ji Hye Hwang1, Soon Hong Yuk, Jin Ho Lee
1Department of Microbiology, The Catholic University of Korea, Seoul 137-701, Korea.
Abstract:
We investigated whether stem cells (MDSC) from primary cultures of rat skeletal muscle can differentiate into the smooth muscle lineage in response to vascular endothelial growth factor (VEGF) and coculture with bladder smooth muscle cells. The MDSC were isolated from gastrocnemius muscle biopsies of normal 3-6 week-old Sprague-Dawley rats and purified by the preplate technique. Cells that took approximately 6 days to adhere to the collagen-coated flasks were termed late preplate (LP) cells, and were used in all the experiments. The early plate (EP) cells (pp1-pp4) contained some myogenic cells but were mostly fibroblasts (< 15% desmin+ cells) whereas the LP cells (pp5-pp6) were highly purified muscle-derived cells (pp6) (> 90% desmin+ cells). The muscle-derived stem cells (LP cells) were CD34+ or Sca-1+, CD45- and desmin+ by immunohistochemical staining. After two days of co-culture with bladder smooth muscle cells, about 25% of the muscle-derived stem cells were positive for alpha-smooth muscle actin (alpha-SMA)+. RT-PCR for alpha-SMA was positive in the VEGF stimulated MDSC, but negative in the absence of VEGF. In conclusion, rat muscle-derived stem cells exhibited stem cell properties (CD34+ or Sca-1+), and were not of hematogeous (CD45-) but of myogenic origin (desmin+). RT-PCR of alpha-SMA was positive in the VEGF stimulated muscle-derived stem cells.
Insights
Rat muscle-derived stem cells (MDSC) can differentiate into smooth muscle cells when stimulated by vascular endothelial growth factor (VEGF). These findings highlight the myogenic potential of MDSC for smooth muscle regeneration.
Area of Science:
- Stem Cell Biology
- Regenerative Medicine
- Muscle Physiology
Background:
- Muscle-derived stem cells (MDSC) are a promising source for regenerative therapies.
- Understanding the differentiation potential of MDSC is crucial for their therapeutic application.
Purpose of the Study:
- To investigate the differentiation capacity of rat skeletal muscle-derived stem cells (MDSC) into the smooth muscle lineage.
- To determine the role of vascular endothelial growth factor (VEGF) and co-culture with bladder smooth muscle cells in this differentiation process.
Main Methods:
- Isolation and purification of late preplate (LP) cells from rat gastrocnemius muscle biopsies.
- Immunohistochemical staining for stem cell markers (CD34, Sca-1, CD45) and myogenic marker (desmin).
- Co-culture of MDSC with bladder smooth muscle cells and stimulation with VEGF, followed by alpha-smooth muscle actin (α-SMA) expression analysis via RT-PCR and immunohistochemistry.
Main Results:
- Late preplate (LP) cells were identified as highly purified muscle-derived stem cells (>90% desmin+).
- These MDSC expressed stem cell markers (CD34+, Sca-1+) and were negative for hematopoietic marker (CD45-).
- Co-culture with bladder smooth muscle cells and VEGF stimulation induced α-SMA expression in approximately 25% of MDSC, indicating smooth muscle differentiation.
Conclusions:
- Rat muscle-derived stem cells possess stem cell properties and are of myogenic origin.
- VEGF stimulation is a key factor in inducing smooth muscle differentiation in MDSC.
- These findings support the potential of MDSC for smooth muscle tissue engineering and regenerative therapies.
