Automated control of cell growth rates for induction and transformation

• US 10,435,662 B1
• Filed: 03/21/2019
• Issued: 10/08/2019
• 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:
  US16/360,404
• Publication Number:
  US 20190300836A1
• Time in Patent Office: 201 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 and devices for automated control of cell growth rates where cell growth is measured in situ and the devices can be used as a stand-alone device or as a module in an automated environment, e.g., as one module in a multi-station or multi-module cell processing environment. The cell growth device comprises a temperature-controlled vial, a motor assembly to spin the vial, a spectrophotometer for measuring, e.g., OD of the cells in the vial, and a processor to accept input from a user and control the growth rate of the cells.

Claims

1. A cell growth device comprising:

a housing;

a motor;

a thermal control device;

a spectrophotometer;

a processor;

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

a rotating growth vial comprising

a vial;

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;

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

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

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

3. The cell growth device of claim 1, wherein optical density is measured at a wavelength selected by a user.

4. The cell growth device of claim 1, wherein the optical density is measured continuously.

5. The cell growth device of claim 1, wherein the optical density is measured at intervals specified by the user or programmed into the processor.

6. The cell growth device of claim 1, wherein the vial volume is 1-250 ml.

7. The cell growth device of claim 6, wherein the vial volume is 2-100 ml.

8. The cell growth device of claim 7, wherein the vial volume is 12-35 ml.

9. The cell growth device of claim 1, wherein the vial is fabricated from polycarbonate or polypropylene.

10. The cell growth device of claim 9, wherein the vial is fabricated by injection molding.

11. The cell growth device 1 of claim 1, further comprising a second light path.

12. The cell growth device of claim 1, wherein the rotating growth vial further ecomprises two or more paddles.

13. The cell growth device of claim 12, wherein the rotating growth vial comprises 2-10 paddles.

14. The cell growth device of claim 13, wherein the rotating growth vial comprises 2-6 paddles.

15. The cell growth device of claim 12, wherein the paddles extend from an inner wall of the rotating growth vial toward the interior of the rotating growth vial.

16. The cell growth device of claim 15, wherein the width of the paddles may range from 1/20 to just under ½

the diameter of the rotating growth vial.

17. The cell growth device of claim 15, wherein the length of the paddles may range from ⅘

to ¼

the length of the main body of the rotating growth vial.

18. The cell growth device of claim 12, wherein the paddles extend from the center of the rotating growth vial toward an inner wall of the rotating growth vial.

19. A cell growth device comprising:

a housing;

a motor;

a thermal control device;

a spectrophotometer;

a processor;

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

a disposable rotating cell growth vial comprising a vial;

a barcode;

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

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

wherein the processor accepts input from a user, receives from the spectrophotometer the optical density of the cells, and through the electrical connection directs the thermal device to adjust the temperature of the vial to grow the cells to a target optical density value at a target time, and wherein the processor is programmed to use wavelength values for blanks commensurate with growth medium in the vial.

20. The cell growth device of claim 19, wherein the processor notifies a user when a desired optical density is reached.