AUTOMATED SYSTEM FOR PRODUCING INDUCED PLURIPOTENT STEM CELLS OR DIFFERENTIATED CELLS

• US 20130345094A1
• Filed: 11/30/2012
• Published: 12/26/2013
• Est. Priority Date: 12/01/2011
• Status: Active Grant

• Current Assignee: New York Stem Cell Foundation
• Original Assignee: New York Stem Cell Foundation
• Inventors: Eggan, Kevin C., Noggle, Scott, Chang, Stephen, Solomon, Susan
• Granted Patent: US 10,273,459 B2
• Time in Patent Office: Days
• US Class Current: 506/14
• 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 iPSCs, comprising:

a cell plating unit for placing cells on a plate; and

an induction unit for automated reprogramming of cells by contacting the cells on the plating unit with reprogramming factors to produce iPSCs.

2. The system of claim 1, further comprising:

a sorting unit for selectively sorting and isolating the iPSCs produced by the induction unit by identifying iPS specific markers.

3. The system of claim 2, further comprising:

an expansion unit for expanding the isolated iPSCs, and selecting the expanded iPSCs.

4. The system of claim 1, further comprising:

a freezing unit for freezing the isolated iPSCs.

5. The system of claim 1, further comprising:

a confluency check unit which checks the confluency of the iPSCs to determine whether the iPSCs have a confluency characteristic.

6. The system of claim 5, wherein the confluency check unit periodically checks the confluency of the iPSCs.

7. The system of claim 5, wherein the confluency check unit periodically checks the confluency of the iPSCs on a more frequent basis over time.

8. The system of claim 5, wherein the confluency check unit checks whether the confluency characteristic is within the range of 70% to 100%.

9. The system of claim 1, wherein the induction unit uses a viral vector to initiate reprogramming.

10. The system of claim 9, wherein the induction unit uses a retrovirus or a Sendai virus to initiate re-programming.

11. The system of claim 1, wherein the induction unit uses small molecules, peptides, proteins or nucleic acids to initiate re-programming.

12. The system of claim 1, further comprising an electroporation unit for electroporating the cells.

13. The system of claim 1, wherein the sorting unit comprises a magnetic sorting unit.

14. The system of claim 1, further comprising a cell banking unit for obtaining cells used by the plating unit.

15. The system of claim 14, wherein the banking unit comprises:

a biopsy plating unit for placing biopsies on a plate;

an outgrowth and passaging unit for growing cells; and

a mycoplasma test unit for testing the presence of mycoplasma.

16. The system of claim 15, wherein the mycoplasma test unit comprises a glow luminescence testing device.

17. The system of claim 15, further comprising:

a distribution unit for distributing expanded cells; and

a storage and retrieval system for storing the cells.

18. The system of claim 17, wherein the storage and retrieval system freezes the cells.

19. The system of claim 3, wherein the expansion unit automatically expands and selects the higher-quality clones.

20. The system of claim 1, wherein the cells are somatic cells.

21. The system of claim 20, wherein the somatic cells are fibroblasts.

47. An induced pluripotent stem cell produced using the system of claim 1.

48. A population of induced pluripotent stem cells produced using the system of claim 1.

49. An array comprising the cells produced from a system of any of claims 1, 22 or 38.

50. The array of claim 49, wherein the array is at least a 6, 24, 96, 384, 1536 sized array

52. A cell bank generated by the system of claim 1, 22 or 38.

53. The system of claim 19, wherein higher-quality clones are consolidated to fewer plates than prior to consolidation.

22. An automated system for generating differentiated adult cells from stem cells comprising:

a stem cell plating unit for placing cells on a plate; and

an induction unit for automated reprogramming of cells by contacting the cells on the stem cell plating unit with reprogramming factors to produce differentiated adult cells.

23. The system of claim 22, further comprising a sorting unit for selectively sorting and isolating the differentiated adult cells produced by the induction unit.

24. The system of claim 23, further comprising:

an expansion unit for expanding the isolated differentiated adult cells, and selecting the expanded differentiated adult cells.

25. The system of claim 24, further comprising:

a freezing unit for freezing the isolated differentiated adult cells.

26. The system of claim 22, wherein the stem cells are iPSCs, embryonic stem cells, hematopoietic stem cells, neural stem cells or mesenchymal stem cells.

27. The system of claim 24, further comprising:

a confluency check unit which checks the confluency of the differentiated adult cells to determine whether the differentiated adult cells have a confluency characteristic.

28. The system of claim 27, wherein the confluency check unit periodically checks the confluency of the differentiated adult cells.

29. The system of claim 22, wherein the induction unit uses a viral vector to initiate reprogramming.

30. The system of claim 29, wherein the induction unit uses a retrovirus or a Sendai virus to initiate re-programming.

31. The system of claim 22, wherein the induction unit uses small molecules, peptides, proteins or nucleic acids to initiate re-programming.

32. The system of claim 22, further comprising an electroporation unit for electroporating the cells.

33. The system of claim 22, wherein the sorting unit comprises a magnetic sorting unit.

34. The system of claim 22, further comprising a banking unit for obtaining cells used by the cell plating unit.

35. The system of claim 34, wherein the banking unit comprises:

a biopsy plating unit for placing biopsies on a plate;

an outgrowth and passaging unit for growing cells; and

a mycoplasma test unit for testing the presence of mycoplasma.

36. The system of claim 35, further comprising:

a distribution unit for distributing expanded differentiated adult cells; and

a storage and retrieval system for storing the differentiated adult cells.

37. The system of claim 36, wherein the storage and retrieval system freezes the cells.

51. The system of claim 22 or 38, wherein the differentiated cells are selected from the group consisting of hematopoetic stem cells, muscle cells, cardiac muscle cells, liver cells, cartilage cells, epithelial cells, urinary tract cells, and neuronal cells.

38. An automated system for generating differentiated adult cells from induced pluripotent stem cells (iPSC), comprising:

an iPSC plating unit for placing iPSCs on a plate; and

an induction unit for automated reprogramming of iPSCs by contacting the iPSCs on the iPSC plating unit with reprogramming factors to produce differentiated adult cells.

39. The system of claim 38, further comprising a sorting unit for selectively sorting and isolating the differentiated adult cells produced by the induction unit by identifying markers specific to the differentiated adult cells.

40. The system of claim 38, further comprising:

an expansion unit for expanding the isolated differentiated adult cells, and selecting the expanded differentiated adult cells.

41. The system of claim 38, further comprising:

a freezing unit for freezing the isolated differentiated adult cells.

42. The system of claim 38, wherein the induction unit uses a viral vector to initiate differentiation.

43. The system of claim 42, wherein the induction unit uses a retrovirus or a Sendai virus to initiate differentiation.

44. The system of claim 38, wherein the induction unit uses small molecules, peptides, proteins or nucleic acids to initiate differentiation.

45. The system of claim 38, further comprising a banking unit for obtaining differentiated cells used by the plating unit.

46. The system of claim 38, wherein the cell banking unit comprises:

a biopsy plating unit for placing biopsies on a plate;

an outgrowth and passaging unit for growing cells; and

a mycoplasma test unit for testing the presence of mycoplasma.