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Evaluation of castor oil-based polyurethane membranes in rat bone-marrow cell culture
Sofia de Amorim Cerejo1, Sheila Canevese Rahal, João Ferreira de Lima Neto
1Advanced Reproduction and Cell Therapy Laboratory, Department of Animal Reproduction and Veterinary Radiology, Faculty of Veterinary Medicine and Animal Science, Sao Paulo State University, Botucatu-SP, Brazil.
Purpose:
To evaluate three methods to isolate rats MSCs and to analyze the potential of a castor oil polyurethane base membrane as a scaffold for MSCs.
Methods:
Four male Wistar rats, aged 20-30 days were used. Bone marrow aspirates from femur and tibia were harvested using DMEM high glucose and heparin. The cell culture was performed in three different ways: direct culture and two types of density gradients. After 15 days, was made the 1st passage and analyzed cell viability with markers Hoerscht 33342 and propidium iodide. The MSCs were characterized by surface markers with the aid of flow cytometry. After this, three types of castor oil polyurethane membranes associated with the MSCs were kept on the 6-well plate for 5 days and were analyzed by optical microscopy to confirm cell aggregation and growth.
Results:
Separation procedures 1 and 2 allowed adequate isolation of MSCs and favored cell growth with the passage being carried out at 70% confluence after 15 days in culture. The cells could not be isolated using procedure 3. When the 3 castor oil polyurethane membrane types were compared it was possible to observe that the growth of MSCs was around 80% in membrane type 3, 20% in type 2, and 10% in type 1.
Conclusion:
Both Ficoll-Hypaque densities allow isolation of rat MSCs, and especially castor oil-based membrane type 3 may be used as a scaffold for MSCs.
Insights
Two density gradient methods effectively isolated rat mesenchymal stem cells (MSCs). Castor oil-based polyurethane membrane type 3 demonstrated significant potential as a scaffold for MSCs, promoting robust cell growth.
Area of Science:
- Biomedical Engineering
- Stem Cell Biology
- Materials Science
Background:
- Mesenchymal stem cells (MSCs) are crucial for regenerative medicine.
- Developing efficient isolation methods and suitable biomaterials is essential for MSC applications.
- Polyurethane membranes offer potential as scaffolds for cell culture and tissue engineering.
Purpose of the Study:
- To compare three distinct methods for isolating rat bone marrow-derived MSCs.
- To assess the efficacy of different castor oil polyurethane membranes as scaffolds for MSCs.
Main Methods:
- Rat bone marrow aspirates were cultured using direct culture and two density gradient methods.
- Cell viability was assessed using Hoerscht 33342 and propidium iodide.
- MSCs were characterized by surface markers via flow cytometry.
- MSC growth and aggregation on three castor oil polyurethane membrane types were analyzed using optical microscopy.
Main Results:
- Density gradient procedures 1 and 2 successfully isolated MSCs, enabling passage at 15 days with 70% confluence.
- Procedure 3 failed to isolate MSCs.
- Castor oil polyurethane membrane type 3 supported approximately 80% MSC growth, significantly outperforming types 2 (20%) and 1 (10%).
Conclusions:
- Ficoll-Hypaque density gradients are effective for isolating rat MSCs.
- Castor oil-based polyurethane membrane type 3 is a promising scaffold for MSC culture and potential therapeutic applications.
