Embryonic stem cells

• US 6,875,607 B1
• Filed: 11/09/1999
• Issued: 04/05/2005
• Est. Priority Date: 11/09/1998
• Status: Expired due to Term

• Current Assignee: ES Cell International Pte Ltd. (Lineage Cell Therapeutics, Inc.)
• Original Assignee: ES Cell International Pte Ltd. (Lineage Cell Therapeutics, Inc.)
• Inventors: Trounson, Alan Osborne, Fong, Chui-Yee, Bongso, Ariffeen, Pera, Martin Frederick, Reubinoff, Benjamin Eithan
• Primary Examiner(s): WOITACH, JOSEPH T
• Application Number:
  US09/436,164
• Time in Patent Office: 1,974 Days
• Field of Search: 435/325, 435/1.1, 800/8
• US Class Current: 435/325
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0652 Cells of skeletal and connective tissues; Mesenchyme

Abstract

The present invention relates to undifferentiated human embryonic stem cells, methods of cultivation and propagation, production of differentiated cells and in particular the production of human embryonic stem cells capable of yielding somatic differentiated cells in vitro, as well as committed progenitor cells capable of giving rise to mature somatic cells and uses thereof. The present invention also provides a purified preparation of undifferentiated human embryonic stem cells capable of proliferation in vitro. Furthermore, the present invention provides a somatic cell differentiated in vitro from an undifferentiated embryonic stem cell. There is also provided a committed progenitor cell capable of giving rise to mature somatic cells.

Claims

1. A method of modulating the differentiation of undifferentiated, pluripotent human embryonic stem (hES) cell in culture, comprising providing a fibroblast feeder layer which has been selected based on its ability to induce differentiation of undifferentiated, pluripotent human embryonic stem (hES) cells in culture, and growing said undifferentiated, pluripotent human embryonic stem (hES) cells on said fibroblast feeder layer, wherein said fibroblast feeder layer modulates the differentiation of said undifferentiated, pluripotent human embryonic stem (hES) cell in culture.

5. The method according to any one of claims 1-4, further comprising cultivating the hES cells for prolonged periods and/or at high density.

6. The method according to any one of claims 1-4, wherein the fibroblast feeder layer is a mouse and/or human fibroblast feeder layer.

7. The method according to any one of claims 1-4, wherein said fibroblast feeder layer comprises embryonic fibroblasts.

8. The method according to any one of claims 1-4, wherein the undifferentiated, pluripotent hES cells are prepared by a process comprising:

obtaining an in vitro fertilised human embryo and growing said embryo to a blastocyst stage of development;

removing inner cells mass (ICM) cells from said embryo;

culturing said ICM cells on the fibroblast feeder layer; and

recovering the ICM cells from the feeder layer as hES cells.

2. A method to of modulating the differentiation of undifferentiated, pluripotent human embryonic stem (hES) cell in culture, comprising providing a fibroblast feeder layer which has been selected based on its ability to favour differentiation of the hES cell into a somatic lineage or into an extraembryonic lineage, and growing said undifferentiated, pluripotent human embryonic stem (hES) cells on said fibroblast feeder layer, wherein said fibroblast feeder layer modulates the differentiation of said undifferentiated, pluripotent human embryonic stem (hES) cell in culture.

3. A method of modulating the differentiation of undifferentiated, pluripotent human embryonic stem (hES) cell in culture, comprising providing a fibroblast feeder layer which has been selected based on its ability to favour differentiation into a somatic lineage and to limit differentiation into an extraembryonic lineage, and growing said undifferentiated, pluripotent human embryonic stem (hES) cells on said fibroblast feeder layer, wherein said fibroblast feeder layer modulates the differentiation of said undifferentiated, pluripotent human embryonic stem (hES) cell in culture.

4. A method of modulating the differentiation of undifferentiated, pluripotent human embryonic stem (hES) cell in culture, comprising providing a fibroblast feeder layer which has been selected based on its ability to induce the differentiation of the hES cell into a somatic lineage or multiple somatic lineages, and growing said undifferentiated, pluripotent human embryonic stem (hES) cells on said fibroblast feeder layer, wherein said fibroblast feeder layer modulates the differentiation of said undifferentiated, pluripotent human embryonic stem (hES) cell in culture.