INCORPORATION OF PHOSPHATIDYLCHOLINE IN A MEDIA COMPOSITION
The present disclosure provides the use of a phosphatidylcholine compound as a component of a media composition. The resulting media composition can be used for the cryopreservation of eukaryotic cells. The cryopreserved eukaryotic cells can be thawed and recovered for inoculation and/or growth in a media to reproduce new cells. Provided herein are compositions and methods for the cryopreservation of eukaryotic cells in a media composition containing a phosphatidylcholine compound.
1 .- 18 . (canceled)
19 . A method for cryopreservation of at least one plant cell in a medium composition, the method comprising the steps of:
(i) suspending the at least one plant cell in an aqueous solution to produce the medium composition, wherein the aqueous solution includes at least one cryopreservation agent, a cell suspension medium, and a phosphatidylcholine, wherein the phosphatidylcholine is added to the medium composition at a concentration from 0.05% to 50% of the total volume of the medium composition;
(ii) acclimating the at least one plant cell and medium composition to a reduced temperature; and,
(iii) cooling the at least one plant cell and medium composition to a temperature of less than 4° C.
20 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the cryopreservation agent is selected from the group consisting of glycerol, DMSO (dimethyl sulfoxide), propylene glycol, ethylene glycol, acetamide, and methanol.
21 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the cryopreservation agent is provided at concentrations from 0.5% to 75% of the total volume of the medium composition.
22 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the cell suspension medium comprises a monocotyledonous plant cell suspension medium.
23 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the cell suspension medium comprises a dicotyledonous plant cell suspension medium.
24 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the plant cell is selected from the group consisting of a monocotyledonous plant cell and a dicotyledonous plant cell.
25 . The method for cryopreservation of at least one plant cell in a medium composition of claim 24 , wherein the monocotyledonous plant cell is selected from the group consisting of maize, wheat, rice, black grass, tall fescue, bent grass, and anza wheat.
26 . The method for cryopreservation of at least one plant cell in a medium composition of claim 24 , wherein the dicotyledonous plant cell is selected from the group consisting of soybean, tobacco, Arabidopsis , canola, cotton, rape, sunflower, and carrot.
27 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the plant cell comprises a transgene.
28 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the phosphatidylcholine is selected from the group consisting of soybean L-α-phosphatidylcholine, rape seed L-α-phosphatidylcholine, cotton seed L-α-phosphatidylcholine, and sunflower seed L-α-phosphatidylcholine.
29 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the medium composition is frozen.
30 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the medium composition reaches a temperature of about 0° C. to about −196° C.
31 . The method for cryopreservation of at least one plant cell in a medium composition of claim 19 , wherein the plant cell is a plant suspension cell line, or a photo-autotrophic suspension cell line, a plant embryo, a plant pollen, meristematic tissue, plant leaf, plant root, plant root tip, anther, silk, flower, kernel, bulb, tuber, rhizome, calli, or seed.