Magnetic extracellular matrix
Methods of making and using a magnetic ECM are disclosed. The ECM comprises positively and negatively charged nanoparticles, wherein one of said nanoparticles contains a magnetically responsive element. When the magnetic ECM is seeded with cells, the cells will be magnetized and can be levitated for 3-D cell culture.
1. A method of making a magnetic extracellular matrix (ECM), said method comprising:
a) combining cells with a composition comprising a magnetic nanoparticle so that said cells take up said magnetic nanoparticle;
b) levitating said cells in a magnetic field;
c) growing said levitated cells in said magnetic field into a 3D cell culture having an ECM until said magnetic nanoparticles are lost from the cells and retained by said ECM; and
d) decellularizing said 3D cell culture using a magnet to hold said ECM while washing away cell debris to produce a magnetic ECM.
2. The method of claim 1 , wherein said magnetic nanoparticles are provided by blasting, injection, electroporation, magnetic pressure, hydrogels, or cationic liposomes.
3. The method of claim 1 , wherein said composition comprises:
i) a negatively charged nanoparticle;
ii) a positively charged nanoparticle; and
iii) a support molecule;
wherein one of said negatively charged nanoparticle or positively charged nanoparticle contains a magnetically responsive element, and wherein said support molecule holds said negatively charged nanoparticle and said positively charged nanoparticle in an intimate admixture.
4. The method of claim 3 , wherein a) the support molecule comprises peptides, polysaccharides, nucleic acids, polymers, poly-lysine, fibronectin, collagen, laminin, BSA, hyaluronan, glycosaminoglycan, anionic, non-sulfated glycosaminoglycan, gelatin, nucleic acid, extracellular matrix protein mixtures, antibody, or mixtures or derivatives thereof, b) said negatively charged nanoparticle is a gold nanoparticle, and c) said positively charged nanoparticle is an iron oxide nanoparticle.
5. The method of claim 1 , further comprising seeding said magnetic ECM with cells, and growing said seeded ECM in a magnetic field.
6. The method of claim 5 , further comprising transplanting the seeded ECM into a patient.
7. The method of claim 1 , further comprising seeding said magnetic ECM with fibroblast and/or epithelial and/or osteoblast cells, plus stem and/or progenitor cells, and further growing said seeded ECM in a magnetic field.
8. The method of claim 1 , wherein said cells are in the form of a tissue fragment.
9. The method of claim 1 , wherein said cells are in the form of a tissue fragment extracted from a patient.
10. The method of claim 9 , wherein the method further comprises seeding said ECM with stem or progenitor cells from said patient, and further growing said seeded ECM in a magnetic field.
11. A method of making a magnetic ECM, said method comprising:
a) placing magnetic nanoparticles inside a cell;
b) levitating said cell in a magnetic field and growing said levitated cell in said magnetic field into a 3D culture of cells having an ECM until said magnetic nanoparticles are lost from said cells and retained by the ECM; and
c) decellularizing said 3D culture of cells using a magnet to levitate and hold said ECM while washing away cell debris to produce a magnetic ECM.