Method of Reseeding Adherent Cells Grown in a Hollow Fiber Bioreactor System
AssigneeTerumo BCT, Inc.
InventorsGlen Delbert ANTWILER, David A. WINDMILLER, Monique GIVENS
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Abstract
This invention is directed to methods of directly reseeding harvested adherent cells grown in a hollow fiber bioreactor. Also disclosed is a novel harvest media for use in directly reseeding adherent cells into a hollow fiber bioreactor.
Claims9
A method for directly reseeding harvested adherent cells grown in growth media in a hollow fiber bioreactor having an intracapillary space and an extracapillary space wherein the cell growth space is one of the intracapillary space or the extracapillary space, comprising the steps of:
- removing the growth media from the cell growth space;
- washing the cells to remove residual growth media from the cell growth space;
- loosening the cells from the cell growth space by adding trypsin to the cell growth space;
- removing the cells and any trypsin from the cell growth space;
- inactivating the trypsin from the removed cells;
- and
- directly reseeding the removed cells and trypsin into the cell growth space of a hollow fiber bioreactor.
A media for harvesting cells grown in a hollow fiber bioreactor comprising;
- base media;
- protein;
- 3.9 mM calcium chloride;
- 9.91 mM anhydrous D-glucose;
- and
- 2.24 mM magnesium sulfide.
A method of making harvest media comprising the steps of;
- calculating the amount of ions and protein needed in the media using Chvh+0vcircuit=cfvh+ccvc;
- and
- adding the calculated amounts of ions and protein to the media.
Dependencies are inferred from the claim text.