Cell culture method
The present invention is directed towards methods for producing cell and tissue compositions suitable for therapeutic applications to a mammal in need of a therapeutic cell or tissue treatment. In particular, the invention is directed towards a method of culturing cells for use in therapy, which method comprises (i) culturing a sample of cells in a first cell culture medium and (ii) prior to harvesting cells for use in therapy culturing the cells in a second cell culture medium.
1. A method of culturing human mesenchymal stromal cells for use in allogeneic therapy comprising the steps of:
(i) providing a sample of said cells from a donor to a first culture medium;
(ii) culturing said cells in the first cell culture medium comprising fetal calf serum, for at least 18 hours until said cells are at least 70% confluent;
(iii) replacing said first cell culture medium with a second cell culture medium comprising about 2% v/v human serum albumin;
(iv) further culturing the cells for between 24 hours and 36 hours in the second cell culture medium; and
(v) harvesting said cells for use in said allogeneic therapy.
2. The method of claim 1 , wherein the mesenchymal stromal cell is a mesenchymal progenitor cell (MPC).
3. The method of claim 1 , wherein the sample of cells in step (i) comprises between 1.0×10 3 cells per cm 2 of culture vessel and 1.0×10 10 cells per cm 2 of culture vessel.
4. The method of claim 1 , wherein the sample of cells in step (i) comprises between 1.0×10 3 cells per cm 2 of culture vessel and 1.0×10 7 cells per cm 2 of culture vessel.
5. The method of claim 1 , wherein the sample of cells in step (i) are cultured until said cells are approximately 85% confluent.
6. The method of claim 1 , wherein the second cell culture medium is phenol red-free.