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Updated: Aug 30, 2026
Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
Published on: February 9, 2015
[Isolation and culture of multipotent mesenchymal stem cells from porcine bone marrow]
Li-jun Fang1, Xiao-bing Fu, Tong-zhu Sun
1Department of Stomatology, the Second Hospital of China Medical University, Shenyang 110003, Liaoning, China.
Objective:
To establish a method for the isolation of porcine mesenchymal stem cells (MSCs) from bone marrow and to demonstrate their differentiation ex vivo into various mesenchymal tissue cells.
Methods:
MSCs were isolated from bone marrow and purified by centrifuge and in vitro. The proliferation and growth characteristics were observed in primary and passage culture. Cell cycle was analyzed by measuring DNA content with FAC-Scan flow cytometer and cell multipotent was identified with specific staining.
Results:
The adherent, fibroblast-like cells were confluent in single layer after plating for 12-14 days. The cultured MSCs in vitro differentiated into osteoblasts. The cell cycle analysis showed that 80% of MSCs were in G0/G1 phase.
Conclusion:
Porcine MSCs can be isolated from postnatal bone marrow through their adherent ability. Porcine MSCs may be introduced as a valuable model system to study the mesenchymal lineages for basic research and tissue engineering.
Insights
Researchers successfully isolated porcine mesenchymal stem cells (MSCs) from bone marrow, demonstrating their potential for differentiation into various cell types for research and tissue engineering applications.
Area of Science:
- Veterinary Science
- Stem Cell Biology
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) are multipotent stromal cells with significant therapeutic potential.
- Porcine models are valuable for translational research due to physiological similarities with humans.
- Establishing reliable methods for porcine MSC isolation is crucial for advancing research.
Purpose of the Study:
- To develop and validate a method for isolating porcine mesenchymal stem cells (MSCs) from bone marrow.
- To confirm the multipotent differentiation capacity of isolated porcine MSCs ex vivo.
- To characterize the growth and cell cycle properties of porcine MSCs.
Main Methods:
- Porcine bone marrow was processed to isolate MSCs using centrifugation and in vitro culture techniques.
- Proliferation and growth kinetics were assessed in primary and passaged cultures.
- Flow cytometry (FAC-Scan) was used for cell cycle analysis, and specific staining identified multipotency.
Main Results:
- Adherent, fibroblastoid cells exhibiting MSC morphology were successfully isolated and cultured.
- In vitro differentiation confirmed the ability of porcine MSCs to differentiate into osteoblasts.
- Cell cycle analysis revealed that approximately 80% of the isolated MSCs were in the G0/G1 phase.
Conclusions:
- Porcine MSCs can be effectively isolated from postnatal bone marrow based on their adherence properties.
- These isolated porcine MSCs represent a promising model for studying mesenchymal lineages.
- The findings support the use of porcine MSCs in basic research and tissue engineering endeavors.