Bag assembly for cultivation of cells

Patent metadata

Field	Value
Publication number	US-11525111-B2
Application number	US-201916586272-A
Country	US
Kind code	B2
Filing date	Sep 27, 2019
Priority date	May 28, 2014
Publication date	Dec 13, 2022
Grant date	Dec 13, 2022

Abstract

The invention discloses a flexible bag assembly for cultivation of cells, comprising one or more bags forming a plurality of cultivation compartments, wherein a drain port in at least a first cultivation compartment is adapted to be fluidically connected with a second cultivation compartment upon opening of a valve means. It also discloses a bioreactor with the bag assembly mounted on a rocking tray and a method of cultivating cells in the assembly.

First claim

The invention claimed is:

1. A flexible multicompartment bag for cultivation of cells in a series of cultivation stages, the bag comprising:
a plurality of serially connected cultivation compartments of different sizes with each compartment in the series adapted for a corresponding stage in the series of cultivation stages respectively; and
a breakable seal between each two adjacent compartments in the series, wherein the breakable seal once opened is configured to fluidically connect the two adjacent compartments upon completion of cultivation of cells in one of the two adjacent compartments such that the cultivated cells can be transferred to the other compartment that is subsequent in the series for subsequent stage of cell cultivation,
wherein the plurality of compartments are provided in a stacked arrangement.

2. The bag of claim 1 , wherein a subsequent cultivation compartment in the series has a volume of at least 120% of the volume of the previous cultivation compartment in the series.

3. The bag of claim 1 , wherein a subsequent cultivation compartment in the series has a volume of at least 150% of the volume of the previous cultivation compartment in the series.

4. The bag of claim 1 , further comprising at least one bag that comprises at least one temperature control compartment.

5. The bag of claim 4 , wherein said temperature control compartment comprises a thermostating fluid inlet and optionally a thermostating fluid outlet.

6. The bag of claim 1 , wherein each cultivation compartment comprises a gas inlet and a gas outlet.

7. The bag of claim 1 , wherein the bag is pre-sterilized prior to cell cultivation.

8. The bag of claim 1 , wherein the bag is pre-sterilized by radiation.

9. A bioreactor assembly, comprising the bag of claim 1 mounted on a tray, adapted to rock back and forth around at least one axis.

10. A method of cultivating cells, comprising the steps of:
a) providing the bioreactor assembly of claim 9 ;
b) introducing culture media and cells in the first cultivation compartment in the series;
c) cultivating the cells in said first cultivation compartment to provide a first cell culture;
d) breaking the breakable seal to fluidically connect said first cultivation compartment with a second cultivation compartment subsequent in the series;
e) transferring said first cell culture to said second cultivation compartment;
f) introducing media to said second cultivation compartment, and;
g) cultivating the cells in said second cultivation compartment to provide a second cell culture.

11. The method of claim 10 further comprising a step e′), after step e), of closing the seal between the first and the second cultivation compartments.

12. The method of claim 10 , further comprising the steps of:
h) breaking a second breakable seal to fluidically connect said second cultivation compartment with a third cultivation compartment subsequent in the series;
i) transferring said second cell culture to said third cultivation compartment;
j) introducing media to said third cultivation compartment, and;
k) cultivating the cells in said third cultivation compartment to provide a third cell culture.

13. The method of claim 12 , further comprising a step i′), after step i), of closing the seal between the second and the third cultivation compartments.

14. A flexible multicompartment bag for cultivation of cells in a series of cultivation stages, the bag comprising:
a plurality of serially connected cultivation compartments of different sizes with at least some of the serially connected compartments in the series adapted for stages in the series of cultivation stages respectively; and
a breakable seal between each two adjacent compartments in the at least some of the serially connected compartments in the series, the breakable seal once opened is configured to fluidically connect the two adjacent compartments upon completion of cultivation of cells in one of the two adjacent compartments such that the cultivated cells can be transferred to the other compartment subsequent in the series for subsequent stage of cell cultivation,
wherein the plurality of compartments are provided in a stacked arrangement.

15. The bag of claim 14 , wherein a subsequent cultivation compartment in the series has a volume of at least 120% of the volume of the previous cultivation compartment in the series.

16. The bag of claim 15 , further comprising at least one bag that comprises at least one temperature control compartment.

17. The bag of claim 16 , wherein said temperature control compartment comprises a thermostating fluid inlet and optionally a thermostating fluid outlet.

18. The bag of claim 14 , wherein at least some of the cultivation compartments comprise a gas inlet and a gas outlet.

19. The bag of claim 14 , wherein the bag is pre-sterilized prior to cell cultivation.

20. A bioreactor assembly, comprising the bag of claim 14 mounted on a tray, adapted to rock back and forth around at least one axis.

21. A method of cultivating cells, comprising the steps of:
a) providing the bioreactor assembly of 20 ;
b) introducing culture media and cells in a first cultivation compartment in the series;
c) cultivating the cells in said first cultivation compartment to provide a first cell culture;
d) breaking a breakable seal to fluidically connect said first cultivation compartment with a second cultivation compartment subsequent in the series;
e) transferring said first cell culture to said second cultivation compartment;
f) introducing media to said second cultivation compartment, and;
g) cultivating the cells in said second cultivation compartment to provide a second cell culture.

22. The method of claim 21 , further comprising a step e′), after step e), of closing the seal between the first and the second cultivation compartments.

23. The method of claim 21 , further comprising the steps of:
h) breaking a second breakable seal to fluidically connect said second cultivation compartment with a third cultivation compartment subsequent in the series;
i) transferring said second cell culture to said third cultivation compartment;
j) introducing media to said third cultivation compartment, and;
k) cultivating the cells in said third cultivation compartment to provide a third cell culture.

24. The method of claim 23 , further comprising a step i′), after step i), of closing the seal between the second and the third cultivation compartments.

Assignees

• Cytiva Sweden Ab

Inventors

• Gebauer Klaus
• Castan Andreas
• Estmer-Nilsson Camilla
• Falkman Thomas
• Wilen Anders
• Miller Michael
• Stankowski Ralph

Classifications

• C12M23/34

  Internal compartments or partitions · CPC title

• C12M23/58

  Reaction vessels connected in series or in parallel (combinations of bioreactors with other apparatus, C12M43/00) · CPC title

• C12M41/24

  inside the vessel · CPC title

• C12M27/16

  Vibrating; Shaking; Tilting · CPC title

• C12M23/14 Primary

  Bags · CPC title