Low rigidity gels for MSC growth modulation
This invention provides gels and matrices having a rigidity in the range of 0.1-2.5 kPa, methods of manufacturing same, and method of preserving a mesenchymal stem cell population or studying mesenchymal stem cells, comprising same.
1. A mesenchymal stem cell (MSC)
isolated from its biological source,
adhered ex vivo onto the surface of a gel or a gel matrix and in the presence of nutrient material, wherein the surface of said gel or said gel matrix has a rigidity of 250 Pa in the microenvironment of the MSC and comprises adhesion molecules, and
in a quiescent state such that the MSC is arrested in the cell cycle;
wherein said quiescent state is characterized by (i) a lack of proliferation, (ii) a lack of differentiation, (iii) the ability of the MSC to express proteins, and (iv) the ability of the MSC to resume proliferation and differentiation upon exposure to a chemical stimulus, a mechanical stimulus, a physical factor or a combination thereof; and
wherein said MSC exhibits a rounded morphology.
2. The mesenchymal stem cell of claim 1 , wherein the MSC is arrested in the G1 phase or the G2 phase.
3. The mesenchymal stem cell of claim 1 , wherein said mesenchymal stem cell is a bone marrow-derived mesenchymal stem cell (MSC), a renal stem cell, a skeletal muscle-derived MSC, bone-derived MSC, a dental pulp MSC, a cardiac muscle-derived MSC, a synovial-fluid derived MSC, an umbilical cord MSC or an adipose-derived MSC.
4. The mesenchymal stem cell of claim 1 , wherein said cell does not incorporate BrdU.
5. The mesenchymal stem cell of claim 1 , wherein said cell does not incorporate Trypan Blue staining.
6. The mesenchymal stem cell of claim 1 , wherein the adhesion molecules comprise collagen and fibronectin.
7. The mesenchymal stem cell of claim 1 , wherein the physical factor is pressure, temperature, or a combination of pressure and temperature.
8. A mesenchymal stem cell (MSC)
isolated from its natural in vivo microenvironment,
adhered onto the surface of a gel or a gel matrix and in the presence of nutrient material, wherein the surface of said gel or said gel matrix has a rigidity of 250 Pa in the microenvironment of the MSC and comprises adhesion molecules, and
in a quiescent state such that the MSC is arrested in the cell cycle;
wherein said quiescent state is characterized by (i) a lack of proliferation, (ii) a lack of differentiation, (iii) the ability of the MSC to express proteins, and (iv) the ability of the MSC to resume proliferation and differentiation upon exposure to a chemical stimulus, a mechanical stimulus, a physical factor, or a combination thereof; and
wherein said cell exhibits a rounded morphology.
9. The mesenchymal stem cell of claim 8 , wherein the MSC is arrested in the G1 phase or the G2 phase.
10. The mesenchymal stem cell of claim 8 , wherein said mesenchymal stem cell is a bone marrow-derived mesenchymal stem cell (MSC), a renal stem cell, a skeletal muscle-derived MSC, bone-derived MSC, a dental pulp MSC, a cardiac muscle-derived MSC, a synovial-fluid derived MSC, an umbilical cord MSC or an adipose-derived MSC.
11. The mesenchymal stem cell of claim 8 , wherein said cell does not incorporate BrdU.
12. The mesenchymal stem cell of claim 8 , wherein said cell does not incorporate Trypan Blue staining.
13. The mesenchymal stem cell of claim 8 , wherein the adhesion molecules comprise collagen and fibronectin.
14. The mesenchymal stem cell of claim 8 , wherein the physical factor is pressure, temperature, or a combination of pressure and temperature.
15. The mesenchymal stem cell of claim 1 , wherein said gel or gel matrix comprises acrylamide and bisacrylamide.
16. The mesenchymal stem cell of claim 8 , wherein said gel or gel matrix comprises acrylamide and bisacrylamide.
17. The mesenchymal stem cell of claim 1 , wherein the adhesion molecules comprise collagen.
18. The mesenchymal stem cell of claim 8 , wherein the adhesion molecules comprise collagen.
19. The mesenchymal stem cell of claim 1 , wherein the adhesion molecules comprise fibronectin.
20. The mesenchymal stem cell of claim 8 , wherein the adhesion molecules comprise fibronectin.