Methods for controlling the growth of prokaryotic and eukaryotic cells

• US 10,443,031 B1
• Filed: 03/21/2019
• Issued: 10/15/2019
• Est. Priority Date: 03/29/2018
• Status: Active Grant

• Current Assignee: Inscripta, Inc.
• Original Assignee: Inscripta, Inc.
• Inventors: Masquelier, Don, Belgrader, Phillip
• Primary Examiner(s): Beisner, William H.
• Application Number:
  US16/360,423
• Publication Number:
  US 20190300840A1
• Time in Patent Office: 208 Days
• CPC Class Codes: C12M 23/08 Flask, bottle or test tube
• CPC Class Codes: C12M 23/44 Multiple separable units; Modules
• CPC Class Codes: C12M 27/02 Stirrer or mobile mixing elements
• CPC Class Codes: C12M 27/14 Rotation or movement of the cells support, e.g. rotated hollow fibers
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• CPC Class Codes: C12M 41/36 of biomass, e.g. colony counters or by turbidity measurements electrooptical investigation of individual particles G01N15/14, flow cytometers G01N15/1404
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 15/87 Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation

Abstract

The present disclosure relates to methods for control of cell growth rates where cell growth is measured in situ. The methods are applicable to bacterial cells, mammalian cells, non-mammalian eukaryotic cells, plant cells, yeast cells, fungi, and archea.

Claims

1. A method for using a cell growth device, comprising:

transferring an aliquot of cells into cell growth device comprising;

a housing;

a motor;

a thermal control device;

an electrical connection configured to be electrically coupled to a thermal control device;

a spectrophotomer;

a processor;

and a rotating growth vial comprising

a vial comprising growth medium;

a drive engagement mechanism configured to engage with the motor to spin the vial; and

a light path through the vial for a light beam generated by the spectrophotometer, wherein the spectrophotometer is configured to measure and deliver to the processor an optical density of cells in the vial;

entering a user-preferred target optical density and a user-preferred time to reach the user-preferred target optical density into the processor wherein the processor accepts input from a user, receives from the spectrophotometer the measure of the optical density of the cells in the vial, and through the electrical connection directs the thermal control device to adjust a temperature of the vial to grow the cells to the user-preferred target optical density at the user-preferred target time; and

wherein the processor is programmed to use wavelength values for blanks commensurate with the growth medium in the vial.

2. The method of claim 1, wherein the cell growth device performs the measuring and adjusting steps until the cells reach the user-preferred optical density of the cells.

3. The method of claim 1, wherein a volume of the cells is from 5-85% of a volume of the rotating growth vial.

4. The method of claim 1, wherein the user-preferred optical density is 2.7.

5. The method of claim 1, wherein the optical density is read at OD₆₀₀.

6. The method of claim 1, wherein the cells are bacterial cells.

7. The method of claim 1, wherein the cells are yeast cells.

8. The method of claim 1, wherein the cells are mammalian cells.

9. The method of claim 8, wherein the mammalian cells are adherent cells.

10. The method of claim 9, wherein the adherent cells are grown on microcarriers.

11. A method for using a cell growth device, comprising:

transferring an aliquot of cells into a cell growth device, wherein the cell growth device comprises;

a housing;

a motor;

a thermal control device;

an electrical connection configured to be electrically coupled to the thermal control device;

a spectrophotometer;

a processor; and

a rotating cell growth vial comprising a medium-filled vial;

a drive engagement mechanism connected to the motor and configured to spin the vial; and

a first light path through the vial to measure an optical density of cells in the vial via the spectrophotometer;

entering a user-preferred target optical density, wherein the processor accepts input from a user and receives from the spectrophotometer the optical density of the cells;

determines when the cells reach the user-preferred target optical density; and

wherein the processor cools the cell growth vial or advances cells to a next module in an automated multi-module cell processing system.

12. The method of claim 11, wherein the cells are bacterial cells.

13. The method of claim 11, wherein the cells are yeast cells.

14. The method of claim 11, wherein the cells are mammalian cells.

15. The method of claim 14, wherein the mammalian cells are adherent cells.

16. The method of claim 15, wherein the adherent cells are grown on microcarriers.

17. The method of claim 11, wherein the cell growth device notifies a user that the cells have reached the user-preferred target optical density.

18. The method of claim 11, wherein the rotating growth vial further comprises a barcode.

19. The method of claim 11, wherein the user-preferred optical density is 2.7.

20. The method of claim 11, wherein the optical density is read at OD₆₀₀.