Search

RU-2861299-C2 - CELL COMPOSITIONS OBTAINED FROM DEDIFERENTIATED REPROGRAMMED CELLS

RU2861299C2RU 2861299 C2RU2861299 C2RU 2861299C2RU-2861299-C2

Abstract

FIELD: biotechnology. SUBSTANCE: invention relates to a composition for obtaining pancreatic hormone-secreting cells, comprising definitive endoderm cells obtained from human induced pluripotent cells, and Y-27632, wherein said human cells exclude cells obtained from human embryonic stem cells obtained exclusively by a method requiring the destruction of an embryo. EFFECT: cell compositions obtained from pluripotent cells, for example, dedifferentiated reprogrammed cells, such as induced pluripotent stem (iPS) cells. 9 cl, 12 tbl, 8 dwg, 7 ex

Inventors

  • AGULNIK, Alan D.
  • KELLI, Oliviya
  • OKHI, Yuki
  • ROBINS, ALLAN
  • SHULTS, Tomas

Dates

Publication Date
20260504
Application Date
20210804
Priority Date
20130206

Claims (9)

  1. 1. A composition for obtaining pancreatic cells that secrete hormones, containing definitive endoderm cells obtained from human induced pluripotent cells and Y-27632, wherein said human cells exclude cells that are obtained from human embryonic stem cells obtained exclusively by a method requiring the destruction of the embryo.
  2. 2. The composition according to claim 1, characterized in that said definitive endoderm cells are capable of differentiating into human pancreatic islet cells that secrete hormones.
  3. 3. The composition according to claim 1, characterized in that said definitive endoderm cells are capable of differentiating into endodermal cells of the anterior part of the human digestive tract.
  4. 4. The composition according to claim 3, characterized in that said definitive endoderm cells are capable of differentiating into PDX1-positive human pancreatic endodermal cells.
  5. 5. The composition according to claim 1, further comprising a growth factor of the Nodal subgroup/activin superfamily TGFβ.
  6. 6. The composition according to claim 5, characterized in that said growth factor of the Nodal/activin subgroup of the TGFβ superfamily is selected from the group consisting of Nodal, activin A, activin B and a combination thereof.
  7. 7. The composition of claim 5, further comprising bone morphogenetic protein (BMP), BMP4, GDF-8, GDF-9, GDF-10 or GDF-11.
  8. 8. The composition of claim 1, further comprising a member of the Wnt family.
  9. 9. The composition according to claim 8, characterized in that said member of the Wnt family is Wnt3A.

Description

Cross-references to related applications This application claims the benefit of U.S. Patent Application No. 13/761,078, entitled "Cell Compositions Derived from Dedifferentiated Reprogrammed Cells," filed February 6, 2013, which is a continuation-in-part of U.S. Patent Application No. 12/765,714, entitled "Cell Compositions Derived from Dedifferentiated Reprogrammed Cells," filed April 22, 2010, which is a non-provisional application claiming priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 61/171,759, entitled "Cell Compositions Derived from Dedifferentiated Reprogrammed Cells," filed April 22, 2009, the disclosure of which is incorporated herein in its entirety by reference. List of sequences This application is filed with an electronic sequence listing. The sequence listing is provided as a file entitled CYTHERA068P1.TXT, created on January 28, 2013, and is 8.92 kb in size. The electronic format of the sequence listing is incorporated herein by reference in its entirety. Fields of technology to which the present invention relates The present invention relates to the isolation, maintenance, and use of cell cultures. In particular, it relates to cell compositions derived from induced pluripotent stem cells. Prior art An important application of pluripotent cells is their use in cell therapy. Pluripotent stem cells include, but are not limited to, human embryonic stem cells (hES) and human embryonic germ cells (hEG). Other types of pluripotent cells exist, such as human and mouse dedifferentiated stem cells, which are differentiated somatic adult cells dedifferentiated to produce pluripotent-like stem cells. These dedifferentiated cells, induced to produce cells with pluripotency and growth capacity similar to ES cells, are also called "induced pluripotent stem (iPS) cells," "embryonic stem cell-like cells," "ES-like cells," or their equivalents. Such cells are potentially viable alternative pluripotent cells. Therapeutic use of iPS cells requires proof that these cells are stable and exhibit a suitable safety profile in preclinical studies for the treatment of diabetes and other diseases. Reprogramming differentiated human somatic cells to a pluripotent state provides patient- and disease-specific stem cells. See Takahashi, K. et al. Cell, 1–12, 2007, and Ju, J. et al. Science 2007. Takahashi et al. and Ju et al. introduced four genes into adult and fetal/neonatal fibroblasts to generate iPS cells: Oct4, Sox2, Klf4, and c-myc in Takahashi et al.; Oct4, Sox2, Nanog, and Lin28 in Ju et al. In each case, iPS cells shared some characteristics of hES cells, including morphology, marker expression, prolonged proliferation, a normal karyotype, and hES cell pluripotency. Although iPS cells may provide a cell-based regenerative tool without associated ethical concerns, the differentiation properties of iPS cells, such as differentiation potential and in vitro differentiation efficiency, remain unclear, and a method for directed differentiation of iPS cells has not been demonstrated. Thus, there is a need to determine and demonstrate the detailed differentiation properties and efficiency of directed differentiation of iPS cells. Statement of the essence of the present invention Embodiments described herein provide cell compositions derived from pluripotent cells, such as dedifferentiated reprogrammed cells such as induced pluripotent stem (iPS) cells. One embodiment provides compositions and methods for producing an in vitro cell culture comprising human cells, wherein at least about 15% of the human cells are definitive endodermal cells, wherein the definitive cells are derived from dedifferentiated, genetically reprogrammed cells. In one aspect, the definitive endodermal cells are multipotent cells that can differentiate into intestinal tube cells or organs derived therefrom. Other embodiments provide compositions and methods for producing an in vitro cell culture comprising human cells, wherein at least about 15% of the human cells are pancreaticoduodenal homeobox factor-1 (PDX1)-positive anterior gastrointestinal tract endodermal cells, wherein the PDX1-positive anterior gastrointestinal tract endodermal cells are derived from dedifferentiated genetically reprogrammed cells. In one aspect, the PDX1-positive anterior gastrointestinal tract endodermal cells are simultaneously PDX1, SOX9, PROX1, and HNF6 positive. Another embodiment provides compositions and methods for producing an in vitro cell culture comprising human cells, wherein at least about 15% of the human cells are pancreatic progenitor cells positive for pancreatic-duodenal homeobox factor-1 (PDX1), wherein the PDX1-positive pancreatic progenitor cells are derived from dedifferentiated genetically reprogrammed cells. In one aspect, the PDX1-positive pancreatic progenitor cells are simultaneously PDX1 and NKX6.1 positive. Another embodiment provides compositions and methods for producing an in vitro cell culture comprising hu