Automated system for producing induced pluripotent stem cells or differentiated cells

• US 10,273,459 B2
• Filed: 11/30/2012
• Issued: 04/30/2019
• Est. Priority Date: 12/01/2011
• Status: Active Grant

• Current Assignee: New York Stem Cell Foundation
• Original Assignee: New York Stem Cell Foundation
• Inventors: Noggle, Scott, Eggan, Kevin C., Chang, Stephen, Solomon, Susan
• Primary Examiner(s): Beisner, William H.
• Assistant Examiner(s): Henkel, Danielle B
• Application Number:
  US13/691,258
• Publication Number:
  US 20130345094A1
• Time in Patent Office: 2,342 Days
• CPC Class Codes: C12M 23/04 Flat or tray type, drawers C12M23/10, C12M23/12, C12M23/16 take precedence
• CPC Class Codes: C12M 35/02 Electrical or electromagnetic means, e.g. for electroporation or for cell fusion
• CPC Class Codes: C12M 35/08 Chemical, biochemical or biological means, e.g. plasma jet, co-culture
• CPC Class Codes: C12M 47/04 Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The invention provides an automated system for producing induced pluripotent stem cells (iPSCs) from adult somatic cells. Further, the system is used for producing differentiated adult cells from stem cells. The invention system is useful for isolating somatic cells from tissue samples, producing iPSC lines from adult differentiated cells by reprogramming such cells, identifying the pluripotent reprogrammed adult cells among other cells, and expanding and screening the identified reprogrammed cells.

Claims

1. An automated system for generating induced pluripotent stem cells (iPSCs), comprising:

an automated cell plating unit for placing cells on a plate;

an automated induction unit configured to control contacting of the cells on the plating unit with reprogramming factors and producing iPSCs;

an automated sorting unit configured to control depleting non-reprogrammed cells from the produced iPSCs, and wherein the automated sorting unit is configured to simultaneously process multiple samples in parallel;

a thawing and infection module comprising a thaw and infection liquid handler, a first automated decapper, a first automated centrifuge, and a first automated microscope;

a sorting module comprising a magnetic sorting unit integrated into a cell sorting liquid handler;

a colony identification module comprising a colony identification liquid handler and a second automated microscope; and

a first incubator and a second incubator;

wherein the liquid handlers, the first automated decapper, the first automated centrifuge, the first automated microscope, and the first incubator are robotically connected;

wherein the second automated microscope and the second incubator are connected to the colony identification liquid handler; and

wherein the automated system comprises controller software having functionality to control automation of the induction unit and the sorting unit.

2. The system of claim 1, wherein the thaw and infection liquid handler comprises a reprogramming factor.

3. The system of claim 2, wherein the reprogramming factor comprises a viral vector.

4. The system of claim 3, wherein the viral vector is a retrovirus or a Sendai virus.

5. The system of claim 2, wherein the reprogramming factor is selected from the group consisting of small molecules, peptides, proteins, and nucleic acids.

6. The system of claim 1, further comprising an electroporation unit.

7. The system of claim 1, further comprising a mycoplasma test unit comprising a quarantine assay liquid handler and an automated plate reader.

8. The system of claim 7, wherein the mycoplasma test unit comprises a glow luminescence testing device.

9. The system of claim 1, further comprising scheduling software.

10. The system of claim 1, wherein the liquid handlers and the second automated microscope are contained within a biosafety cabinet.

11. The system of claim 1, wherein the first automated decapper, the first automated centrifuge, and the first automated microscope are enclosed in a Hepa-filtered hood.

12. The system of claim 1, further comprising:

a maintenance module comprising an expansion and passage liquid handler, a second automated decapper, a second automated centrifuge, a third automated microscope, and an automated plate sealer; and

a third incubator;

wherein the expansion and passage liquid handler, the second automated decapper, the second automated centrifuge, the third automated microscope, the automated plate sealer, and the third incubator are connected to a robotic transport rail.

13. The system of claim 12, further comprising a quality control (QC) module comprising a QC liquid handler, wherein the QC liquid handler is connected to the robotic transport rail.

14. The system of claim 12, further comprising a freezing module comprising a freezing liquid handler and an automated freezer, wherein the freezing liquid handler is connected to the robotic transport rail.

15. The system of claim 12, further comprising a fourth incubator, wherein the fourth incubator is connected to the robotic transport rail.

16. The system of claim 12, further comprising scheduling software.

17. The system of claim 12, wherein the expansion and passage liquid handler is contained within a biosafety cabinet.

18. The system of claim 12, wherein the second automated decapper, the second automated centrifuge, the third automated microscope, and the automated plate sealer are enclosed in a Hepa-filtered hood.

19. The system of claim 13, wherein the QC liquid handler is contained within a biosafety cabinet.

20. The system of claim 14, wherein the freezing liquid handler is contained within a biosafety cabinet.

30. The system of claim 1, wherein the automated induction unit is further configured to control contacting of the cells on the plating unit with differentiation factors and producing differentiated cells.

21. An automated system for generating induced pluripotent stem cells (iPSCs), comprising:

an automated cell plating unit for placing cells on a plate;

an automated induction unit configured to control contacting of the cells on the plating unit with reprogramming factors and producing iPSCs;

an automated sorting unit configured to control depleting non-reprogrammed cells from the produced iPSCs, and wherein the automated sorting unit is configured to simultaneously process multiple samples in parallel;

a thawing and infection module comprising a thaw and infection liquid handler, a first automated decapper, a first automated centrifuge, and a first automated microscope;

a sorting module comprising a magnetic sorting unit integrated into a cell sorting liquid handler;

a colony identification module comprising a colony identification liquid handler and a second automated microscope;

a maintenance module comprising an expansion and passage liquid handler, a second automated decapper, a second automated centrifuge, a third automated microscope, and an automated plate sealer;

a quality control (QC) module comprising a QC liquid handler;

a freezing module comprising a freezing liquid handler and an automated freezer; and

a first incubator, a second incubator, a third incubator, and a fourth incubator;

wherein the thaw and infection liquid handler, the first automated decapper, the first automated centrifuge, the first automated microscope, the cell sorting liquid handler, the colony identification liquid handler, the first incubator, the expansion and passage liquid handler, the second automated decapper, the second automated centrifuge, the third automated microscope, the automated plate sealer, the QC liquid handler, the freezing liquid handler, the third incubator, and the fourth incubator are robotically connected;

wherein the second automated microscope and the second incubator are connected to the colony identification liquid handler; and

wherein the automated system comprises controller software having functionality to control automation of the induction unit and the sorting unit.

22. The system of claim 21, wherein the thaw and infection liquid handler comprises a reprogramming factor.

23. The system of claim 22, wherein the reprogramming factor comprises a viral vector.

24. The system of claim 23, wherein the viral vector is a retrovirus or a Sendai virus.

25. The system of claim 22, wherein the reprogramming factor is selected from the group consisting of small molecules, peptides, proteins, and nucleic acids.

26. The system of claim 21, further comprising an electroporation unit.

27. The system of claim 21, further comprising a mycoplasma test unit comprising a quarantine assay liquid handler and an automated plate reader.

28. The system of claim 27, wherein the mycoplasma test unit comprises a glow luminescence testing device.

29. The system of claim 21, further comprising scheduling software.

31. The system of claim 21, wherein the automated induction unit is further configured to control contacting of the cells on the plating unit with differentiation factors and producing differentiated cells.

32. An automated system for generating cells, comprising:

an automated cell plating unit for placing cells on a plate;

an automated induction unit configured to control contacting of the cells on the plating unit with reprogramming or differentiation factors;

an automated sorting unit configured to control sorting of the cells produced by the automated induction unit, and wherein the automated sorting unit is configured to simultaneously process multiple samples in parallel;

a thawing and infection module comprising a thaw and infection liquid handler, a first automated decapper, a first automated centrifuge, and a first automated microscope;

a sorting module comprising a magnetic sorting unit integrated into a cell sorting liquid handler;

a colony identification module comprising a colony identification liquid handler and a second automated microscope; and

a first incubator and a second incubator;

wherein the liquid handlers, the first automated decapper, the first automated centrifuge, the first automated microscope, and the first incubator are robotically connected;

wherein the second automated microscope and the second incubator are connected to the colony identification liquid handler; and

wherein the automated system comprises controller software having functionality to control automation of the induction unit and the sorting unit.

33. An automated system for generating cells, comprising:

an automated cell plating unit for placing cells on a plate;

an automated induction unit configured to control contacting of the cells on the plating unit with reprogramming or differentiation factors to produce reprogrammed or differentiated cells;

an automated sorting unit configured to control sorting of the cells produced by the automated induction unit, and wherein the automated sorting unit is configured to simultaneously process multiple samples in parallel;

a thawing and infection module comprising a thaw and infection liquid handler, a first automated decapper, a first automated centrifuge, and a first automated microscope;

a sorting module comprising a magnetic sorting unit integrated into a cell sorting liquid handler;

a colony identification module comprising a colony identification liquid handler and a second automated microscope;

a maintenance module comprising an expansion and passage liquid handler, a second automated decapper, a second automated centrifuge, a third automated microscope, and an automated plate sealer;

a quality control (QC) module comprising a QC liquid handler;

a freezing module comprising a freezing liquid handler and an automated freezer; and

a first incubator, a second incubator, a third incubator, and a fourth incubator;

wherein the thaw and infection liquid handler, the first automated decapper, the first automated centrifuge, the first automated microscope, the cell sorting liquid handler, the colony identification liquid handler, the first incubator, the expansion and passage liquid handler, the second automated decapper, the second automated centrifuge, the third automated microscope, the automated plate sealer, the QC liquid handler, the freezing liquid handler, the third incubator, and the fourth incubator are robotically connected;

wherein the second automated microscope and the second incubator are connected to the colony identification liquid handler; and

wherein the automated system comprises controller software having functionality to control automation of the induction unit and the sorting unit.