Cell cultures comprising poly(oxazoline) stabilizers and use of poly(oxazolines) for stabilizing cell cultures

Patent metadata

Field	Value
Publication number	US-2020339945-A1
Application number	US-201816616727-A
Country	US
Kind code	A1
Filing date	May 28, 2018
Priority date	May 26, 2017
Publication date	Oct 29, 2020
Grant date	—

Abstract

Cell cultures comprising poly(oxazoline) stabilizers and use of poly-(oxazolines for stabilizing cell cultures
Described are cell cultures containing in a cell culture medium one or more water-soluble poly(oxazolines).
The water-soluble poly(oxazoline) acts as a stabilizer for the cells and reduces the mechanical stress exerted by agitating the cell culture medium on the cells. This leads to an improved survival rate of the cells compared to the unstabilized state.

First claim

1 . Cell cultures comprising one or more water-soluble poly(oxazolines) in a cell culture medium, provided that the cell culture medium contains essentially no serum.

2 . Cell cultures according to claim 1 , characterized in that the cell culture medium is essentially free of animal components.

3 . Cell cultures according to claim 1 , characterized in that the used cell culture medium is serum-free and/or protein-free.

4 . Cell cultures according to claim 1 , characterized in that the cell culture medium is a chemically defined medium.

5 . Cell cultures according to claim 1 , characterized in that these contain cells, which are suspended in the cell culture medium.

6 . Cell cultures according to claim 1 , characterized in that the cells are suspensions-adapted cells.

7 . Cell cultures according to claim 1 , characterized in that these are used for protein production, virus and/or virus particle production, investigation of metabolism, division and/or other cellular processes.

8 . Cell cultures according to claim 1 , characterized in that these contain cell lines selected from at least one of the following groups: 293-T, A431, A549, BCP1, bEnd.3, BHK-21, BxPC-3, BY-2, CHO, CMT, COS-1, COS-7, CV-1, EPC, HDMEC-T, HEK, HeLa, HepG2, HL-60, HMEC-1, HUVEC-T, HT-1080, Jurkat, K562, LNCaP, MCF-7, MCF-10A, MDCK II, MDT-1A, MyEnd, Neuro-2A, NIH-3T3-T, NTERA-2 cl.D1, P19, PANC-1, Peer, RTL-W1-T, Sf-9, Saos-2, T2, T84 or U-937.

9 . Cell cultures according to claim 8 , characterized in that the cell lines are selected from the group consisting of CHO or HEK, in particular HEK 293.

10 . Cell cultures according to claim 1 , characterized in that these contain hybridoma cells.

11 . Cell cultures according to claim 1 , characterized in that these contain stem cells.

12 . Cell cultures according to claim 1 , characterized in that these contain 0.01 to 15 wt. %, preferably 0.075 to 15 wt. %, more preferred 0.05 to 10 wt. %, highly preferred 0.1 to 10 wt. % of water-soluble poly(oxazolines) in relation to the total amount of cell culture.

13 . Cell cultures according to claim 1 , characterized in that the water-soluble poly(oxazoline) contains at least 80 wt. %, based on its total mass, of repeat structural units of formula I and/or formula II
(I), —NR 1 —CH 2 —CH 2 —CH 2 —  (II),
in which R 1 means a residue of formula —CO—R 2 , R 2 is selected from the group consisting of hydrogen, methyl, ethyl, —C m H 2m —X or —(C n H 2n —O) o -(C p H 2p —O) q —R 3 , R 3 is hydrogen or C 1 —C 6 -alkyl, m is an integer from 1 to 6, X is selected from the group consisting of hydroxyl, alkoxy, amino, N-alkylamino, N,N-dialkylamino, carboxyl, carboxylic ester, sulfonyl, sulfonic acid ester or carbamate, n and p independently of one another are integers from 2 to 4, wherein n is unlike p, and o and q independently of one another are integers from 0 to 60, wherein at least one of o or q is unlike 0.

14 . Cell cultures according to claim 13 , characterized in that the water-soluble poly(oxazoline) has at least 90 wt.-%, based on its total mass, of repeat structural units of formula I, wherein R 2 means methyl or ethyl.

15 . Cell cultures according claim 1 , characterized in that these contain one or more active ingredients, in particular pharmaceutically active ingredients and/or vaccines.

16 . A method for protein production, virus and/or viral particle production, investigation of metabolism, division and/or other cellular processes comprising the cultivation of cell cultures comprising one or more water-soluble poly(oxazolines) in a cell culture medium, provided that the cell culture medium contains essentially no serum.

17 . The method according to claim 16 , characterised in that it also contains the following measures:
(i) filling a bioreactor with one of the cell cultures, and (ii) agitating the contents of the bioreactor.

18 . The method according to claim 16 , characterized in that the duration of the cultivation of the cells in the case of a batchwise method is 5 hours to 30 days, preferably 5 hours to 21 days and in particular 10 hours to 15 days and in the case of a continuous process is 10 days to 180 days, preferably 10 days to 60 days and in particular 15 days to 35 days.

19 . The method according to claim 17 , characterized in that the content of the bioreactor is stirred.

20 . The method according to claim 17 , characterized in that the bioreactor contains baffles for the generation of turbulence.

21 . (canceled)

22 . (canceled)

23 . (canceled)

24 . (canceled)

25 . (canceled)

Assignees

• Univ Jena Friedrich Schiller

Inventors

• Leiske Meike Nicole
• Träger Anja
• Schubert Ulrich Sigmar

Classifications

• C12N2500/50

  Soluble polymers, e.g. polyethyleneglycol [PEG] · CPC title

• C12M27/02

  Stirrer or mobile mixing elements · CPC title

• C12M27/20

  Baffles; Ribs; Ribbons; Auger vanes · CPC title

• C12N5/0018 Primary

  Culture media for cell or tissue culture (media for specific animal cell type C12N5/06) · CPC title

• C12N5/0031 Primary

  Serum-free culture media · CPC title