Deep eutectic solvents and/or ionic liquids in cell culture media

Abstract

The present invention relates to cell culture media compositions comprising deep eutectic solvents and/or ionic liquids.

Claims

1. A cell culture medium kit, comprising, in said kit separately, a dry powder cell culture medium component and a liquid cell culture medium component which are both used to form a composition, wherein the liquid cell culture medium component comprises a deep eutectic solvent comprising choline and/or betaine and/or a derivative or salt thereof and one or more amino acids, and wherein the deep eutectic solvent does not comprise more than 50% water based on weight.

2. The cell culture medium kit according to claim 1, wherein the composition is a base medium or a feed medium.

3. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component is liquid at or below 35 C.

4. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component comprises choline and/or betaine or a salt thereof.

5. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component comprises a quaternary ammonium salt.

6. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component comprises choline and betaine.

7. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component comprises cysteine and/or tyrosine.

8. The cell culture medium kit according to claim 1, wherein the liquid cell culture medium component comprises ingredients which are not part of the deep eutectic solvent and which are dissolved in the deep eutectic solvent.

9. The cell culture medium kit according to claim 1, wherein the deep eutectic solvent does not comprise more than 10% water based on weight.

10. A process for cell culture, comprising: mixing the composition from the cell culture medium kit according to claim 1 with water or with a composition containing water, and adding the resultant composition to a bioreactor either as a base medium to which cells are added afterwards to start a cell culture or as a feed medium to a cell culture already present in the bioreactor.

11. The process for cell culture according to claim 10, wherein the liquid cell culture medium component is liquid at a temperature between 20 and 35 C.

12. The process for cell culture according to claim 10, wherein the liquid cell culture medium component comprises cysteine and/or tyrosine.

13. The process for cell culture according to claim 10, wherein the cells in to the bioreactor are stem cells, eukaryotic cells, prokaryotic cells, archaea, bacteria, yeasts, fungi, insect cells or algae.

14. The process for cell culture according to claim 10, wherein the liquid cell culture medium component comprises less than 10% of water.

15. A cell culture medium, comprising a composition that has been formed from a dry powder cell culture medium component and a liquid cell culture medium component, wherein the liquid cell culture medium component comprises a deep eutectic solvent comprising choline and/or betaine and/or a derivative or salt thereof and one or more amino acids, and wherein the deep eutectic solvent does not comprise more than 50% water based on weight.

16. The cell culture medium according to claim 15, wherein the deep eutectic solvent does not comprise more than 10% water based on weight.

17. The cell culture medium according to claim 15, which comprises choline and/or betaine or a salt thereof; and/or a quaternary ammonium salt; and/or cysteine and/or tyrosine.

Description

EXAMPLES

(1) Comparison of Dissolution Time

(2) The dissolution time of a dry powder tyrosine HCl shall be compared with the dissolution time of a DES comprising Tyrosine HCl. The final concentration of tyrosine in the resulting liquid is set with 0.26 g/L.

(3) Procedure:

(4) A stirrer tank (280 rpm) at 37 C. is filled with 800 ml of a 100 mM HEPES buffer, the pH is adjusted to pH 7 with 2 mol/l NaOH and then the tyrosine sample (powder or DES) is poured onto the surface of the buffer solution.

(5) DES: Tyr HCl:ChCl (1:3), 0.79 g

(6) Powder: a powder mixture of 0.208 g Tyr HCl and 0.36 g ChCl

(7) Focused Beam Reflectance Measurement-Sensor

(8) Mettler Toledo G400

(9) Software: iC FBRM 4.3

(10) FIGS. 1 to 5 show the dissolution of the dry powder and the DES respectively. The x-axis shows the time in all Figures.

(11) A comparison of FIGS. 1 and 2 shows that in case of the DES (FIG. 2), almost no particles or disturbance can be detected in the mixture and complete dissolution is reached after about 10 minutes, whereas in case of the powder (FIG. 1), dissolution takes about 1 hour or more.

(12) The same can also be seen from FIGS. 3 to 5 which show the same measurement in more detail. FIG. 3 is directed to the change of particle number over the time. FIGS. 4 and 5 show the content of particle of different sizes in solution over time.

BRIEF DESCRIPTION OF DRAWINGS

(13) FIG. 1 shows a comparison between the dissolution of a dry powder tyrosine HCl sample and a DES comprising tyrosine HCl.

(14) FIG. 2 shows a comparison between the dissolution of a dry powder tyrosine HCl sample and a DES comprising tyrosine HCl.

(15) FIG. 3A shows the dissolution of a dry powder tyrosine HCl sample.

(16) FIG. 3B shows the dissolution of a DES comprising tyrosine HCl.

(17) FIG. 4 shows a comparison between the dissolution of a dry powder tyrosine HCl sample and a DES comprising tyrosine HCl.

(18) FIG. 5 shows a comparison between the dissolution of a dry powder tyrosine HCl sample and a DES comprising tyrosine HCl.