Methods of modifying transcriptional regulatory networks in stem cells

• US 9,732,128 B2
• Filed: 10/21/2011
• Issued: 08/15/2017
• Est. Priority Date: 10/22/2010
• Status: Active Grant

• Current Assignee: Lineage Cell Therapeutics, Inc.
• Original Assignee: Biotime, Inc.
• Inventors: Chapman, Karen B., West, Michael D.
• Primary Examiner(s): Babic, Christopher M
• Assistant Examiner(s): Aron, Kimberly A
• Application Number:
  US13/279,123
• Publication Number:
  US 20120129262A1
• Time in Patent Office: 2,125 Days
• Field of Search: None
• CPC Class Codes: C07K 14/4702 Regulators; Modulating activity
• CPC Class Codes: C12N 2501/602 Sox-2
• CPC Class Codes: C12N 2501/603 Oct-3/4
• CPC Class Codes: C12N 5/0696 Artificially induced pluripotent stem cells, e.g. iPS

Abstract

The vast differentiation potential of human embryonic and induced pluripotent stem cells, including their potential to cascade through all of the somatic cell lineages and to display the complete transcriptional regulatory network of human biology, has generated interest in deriving scalable, purified, and identified cell types and methods of discovering the precise structure of the human regulatory network. However, the innate capacity of pluripotent cells to display all these lineages is not necessarily reflected during their culture in vitro. The clonal isolation and propagation of progenitors greatly facilitates the generation of highly purified and identified formulations for research and therapeutic purposes. Nevertheless, other cell types have yet to be isolated and propagated from normal cells and methods of isolating said novel cell types as well as methods for introducing perturbations into the transcriptional regulatory network in order to construct a computer model of the entire human transcriptional regulatory network would greatly benefit basic research as well as manufacturing technology for cell-based therapies.

Claims

1. An isolated progenitor cell line, wherein said isolated progenitor cell line constitutively expresses OCT4 and a transcription factor selected from the group consisting of SIX1, FOXA1, SOX17, SIX2, SOX21, PAX6, MYOD1, MYOG, NEUROG1, NKX2.5, and combinations thereof, wherein the expression level of OCT4 is greater than the expression level of OCT4 in an iPS cell.

2. The isolated progenitor cell line of claim 1, wherein said progenitor cell line expresses LHX3.

3. An isolated progenitor cell line, wherein said progenitor cell line expresses the genes RESP18 and LHX3 simultaneously and is the in vitro progeny of a pluripotent stem cell.

5. The isolated progenitor cell line of claim 3, wherein the cell line expresses the gene ESR2.

4. An isolated progenitor cell line, wherein said isolated progenitor cell line constitutively expresses a transcription factor and is a differentiated cell line differentiated in vitro from a pluripotent stem cell.