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 magnetic extracellular matrix (ECM) made by:
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.
2. The magnetic ECM of claim 1 , wherein said magnetic nanoparticles further comprise:
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 mat-like admixture.
3. The magnetic ECM of claim 1 , wherein decellularizing step consists essentially of one or more of freeze-thaw, detergent, or osmotic pressure treatments applied to said 3D culture of cells.
4. The magnetic ECM of claim 2 , wherein decellularizing step consists essentially of one or more of freeze-thaw, detergent, or osmotic pressure treatments applied to said 3D culture of cells.
5. A magnetic extracellular matrix (ECM) made by:
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, wherein said decellularizing consists essentially of one or more of freeze-thaw, detergent, or osmotic pressure treatments applied to said 3D culture of cells.
6. A method of preparing tissue for implantation in a patient, said method comprising:
a) seeding the magnetic ECM of claim 1 with allogeneic stem or progenitor cells from a patient;
b) growing said seeded ECM in a magnetic field; and
c) transplanting the seeded ECM from step b) into said patient.
7. The method according to claim 6 , wherein in step a) the magnetic ECM is further seeded with fibroblast cells, epithelial cells, osteoblast cells, or autologous fibroblast cells and epithelial cells.
8. A method of preparing a seeded magnetic ECM, said method comprising:
a) seeding the magnetic ECM of claim 1 with allogeneic stem or progenitor cells from a patient;
b) growing said seeded ECM in a magnetic field.
9. The method according to claim 8 , wherein in step a) the magnetic ECM is further seeded with fibroblast cells, epithelial cells, osteoblast cells or autologous fibroblast cells and epithelial cells.
10. A seeded magnetic ECM made by the method of claim 8 .