Related Experiment Video
Updated: Jul 15, 2026
Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats
Published on: August 25, 2020
[Isolation and culturation, phenotype detection of rat bone marrow mesenchymal stem cells]
Chao Wang1, Yun Xu, Wen-gang Song
1College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Taian 271018, China. wchxy1617@163.com
Aim:
To investigate the more effective methods of isolation, culture of mesenchymal stem cell (MSC), and examine the surface phenotype of MSC. MSC were separated from rats bone marrow by plastic adherence methods and purified by controlling the time of digestion combined with adhesion separation, and proliferated in culture flasks that were coated with I collogen.
Methods:
The morphology of MSC were studied with phase contrast microscope; cell cycles of the forth generation of MSC were tested by flow cytometry and the phenotype of MSC were identified by using immunocytochemical methods.
Results:
Primary cultured MSC were oval, spindle-shaped or polygonal, and adhered to plastic surface within 24 h and reached 90% confluence within 7-8 days. After purification and proliferation, they were uniformly long spindle-shaped form and passaged every five days. The adhesion rate within 24 hours was all. The flow cytometry showed 80% cells of the forth generation of MSC were at G0/G1 phase. Immunocytochemistry showed MSC were positive for CD29, CD105, CD166, VLA-4 and P-selectin, while negative for CD34 and CD45.
Conclusion:
The MSC obtained from our experiments were more purified, and expanded more rapidly, expressed some cell adhesion molecules. The protocol should make it possible to undertake a large number of experiments with MSC in future application.
Insights
This study optimized mesenchymal stem cell (MSC) isolation and culture, yielding highly purified MSC with rapid expansion and specific cell adhesion molecule expression for future research applications.
Area of Science:
- Stem Cell Biology
- Biotechnology
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) are crucial for tissue repair and regeneration.
- Efficient isolation and culture methods are vital for their therapeutic applications.
- Characterizing MSC surface phenotype is essential for quality control.
Purpose of the Study:
- To optimize mesenchymal stem cell (MSC) isolation and culture techniques.
- To examine the surface phenotype of cultured MSCs.
- To establish a reliable protocol for large-scale MSC expansion.
Main Methods:
- Rat bone marrow MSCs isolated using plastic adherence and controlled digestion.
- Proliferation in collagen-coated flasks.
- Morphological analysis via phase-contrast microscopy.
- Cell cycle analysis by flow cytometry.
- Immunocytochemical identification of surface markers.
Main Results:
- Optimized protocol yielded purified, rapidly expanding MSCs with uniform spindle shape.
- MSCs exhibited high G0/G1 phase cell cycle distribution (80%).
- Confirmed MSC phenotype: positive for CD29, CD105, CD166, VLA-4, P-selectin; negative for CD34, CD45.
Conclusions:
- The developed protocol enhances MSC purification and proliferation rates.
- Cultured MSCs express key cell adhesion molecules, indicating purity and functionality.
- This method facilitates large-scale MSC production for future research and therapeutic use.
