Systems and methods for producing stem cells and differentiated cells

• US 10,428,309 B2
• Filed: 06/27/2014
• Issued: 10/01/2019
• 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, Chang, Stephen, Solomon, Susan
• Primary Examiner(s): Beisner, William H.
• Assistant Examiner(s): Henkel, Danielle B
• Application Number:
  US14/318,339
• Publication Number:
  US 20150166964A1
• Time in Patent Office: 1,922 Days
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• 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 present invention provides various improved systems and methods for obtaining, generating, culturing, and handling cells, such as stem cells (including induced pluripotent stem cells or iPSCs) and differentiated cells, as well as cells and cell panels produced using such systems and methods, and uses of such cells and cell panels.

Claims

1. A method for the automated generation and/or manipulation of stem cells, the method comprising:

a) providing an automated system having controller software,

b) culturing multiple different samples of cells, wherein the cells comprise adult somatic cells,

c) determining the proliferation rate or cell doubling time of individual cell samples from among the multiple different samples of cells utilizing an automated device having controller software for determining the proliferation rate or cell doubling time,

d) freezing individual cell samples from among the multiple different samples of cells,

e) selecting from the individual cell samples a subset of samples having similar proliferation rates or cell doubling times,

f) thawing the subset of samples selected in step (e),

g) plating the subset of samples in a multi-well plate,

h) culturing the subset of samples until they reach a desired confluency,

i) contacting the cultured subset of samples with one or more reprogramming factors in order to produce iPSCs, and

j) sorting the iPSCs and consolidating the sorted iPSCs to produce a pooled iPSC sample, wherein (a) to (j) are performed on the automated system via the controller software.

2. The method of claim 1, wherein the cells are derived from human donors.

3. The method of claim 1, wherein the adult somatic cells are fibroblasts.

4. The method of claim 1, wherein the individual cell samples in the subset of samples have proliferation rates or cell doubling times that are within 10% of one another.

5. The method of claim 1, wherein the culturing of step (h) is performed in low serum medium.

6. The method of claim 1, wherein the step of selecting a subset of samples further comprises selecting samples based the age, sex, race, ethnicity, diagnosis, genotype, phenotype, blood type, HLA type, treatment history, or drug response profile of the cell sample or of the donor from which the cell sample was obtained.

7. The method of claim 1, comprising contacting the pooled iPSCs with one or more differentiation factors thereby producing differentiated cells.

8. The method of claim 1, comprising generating embryoid bodies (EBs) from the pooled iPSCs.

9. The method of claim 8, wherein generating embryoid bodies (EBs) comprises plating iPSCs in V-bottom plates.