Methods and systems for harvesting cells
Methods for using vibration to harvest cells grown in 3D culture are provided. The methods entail the application of force cells attached to a 3D matrix of sufficient amplitude, frequency, and duration to detach cells from the matrix and to flush the detached cells out of the matrix material. An apparatus for performing the methods of the invention as provided.
1. A method of seeding cells in a 3-dimensional matrix in a chamber of a bioreactor, comprising:
a) providing a three-dimensional matrix in a fluid within a container, which container is disposed within said chamber;
b) introducing a composition comprising cells into the container;
c) vibrating the container in a reciprocating motion relative to said chamber for a period of time at a frequency and amplitude sufficient to mix the cells throughout the matrix; and
d) discontinuing the vibration to permit the cells to attach to the matrix.
2. The method of claim 1 , wherein the fluid is a growth medium.
3. The method of claim 2 , further comprising growing the cells in the bioreactor by applying intermittent vibration to the container for a period of time at a frequency and amplitude sufficient to mix the growth medium throughout the matrix.
4. The method of claim 1 , wherein the 3-dimensional matrix is enclosed in a packed bed within said container.
5. The method of claim 4 , wherein the 3-dimensional matrix comprises a single-piece scaffold, multiple beads, multiple carriers, microfibers, nanofibers, or combinations thereof.
6. The method of claim 5 , wherein the microfibers or nanofibers are woven or non-woven.
7. The method of claim 5 , wherein the beads are porous.
8. The method of claim 1 , wherein the 3-dimensional matrix comprises an adherent material.
9. The method of claim 8 , where the adherent material comprises one or more of a polyester, a polypropylene, a polyalkylene, a polyfluorochloroethylene, a polyvinyl chloride, a polyvinyl fluoride resin, a polystyrene, a polysulfone, a polyurethane, a polyethyene terephtalate, a cellulose, a glass fiber, a ceramic particle, a matrigel, an extracellular matrix component, a collagen, a poly L lactic acid, a dextran, an inert metal fiber, silica, natron glass, borosilicate glass, chitosan, or a vegetable sponge.
10. The method of claim 8 , wherein the adherent material is electrostatically charged.
11. The method of claim 1 , wherein the cells are human cells.
12. The method of claim 11 , wherein the human cells are adherent cells.
13. The method of claim 12 , wherein the adherent cells are adherent stromal cells.
14. The method of claim 13 , wherein the origin of the adherent stromal cells is placenta, adipose tissue, or bone marrow.
15. The method of claim 14 , wherein the origin of the adherent stromal cells is placenta.
16. The method of claim 15 , wherein the adherent stromal cells are obtained from a fetal part of the placenta.
17. The method of claim 15 , wherein the adherent stromal cells are obtained from a maternal part of the placenta.
18. The method of claim 1 , wherein the container is vibrated by a substantially linear reciprocating motion.
19. The method of claim 18 , wherein the reciprocating motion has an amplitude of between about 10 mm to about 750 mm and a frequency of 1 to 3 Hz.
20. The method of claim 18 , wherein the amplitude of the substantially linear reciprocating motion is a distance that is 15-100% of the height of said container.