Culturing human embryonic stem cells with a noggin to generate cells lacking Pax-6 expression

• US 9,080,147 B2
• Filed: 01/30/2009
• Issued: 07/14/2015
• Est. Priority Date: 06/20/2000
• 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: Pera, Martin Frederick
• Primary Examiner(s): Crouch, Deborah
• Assistant Examiner(s): SGAGIAS, MAGDALENE K
• Application Number:
  US12/363,194
• Publication Number:
  US 20090197334A1
• Time in Patent Office: 2,356 Days
• US Class Current: 1/1
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/33 Insulin
• CPC Class Codes: C12N 2502/088 neural stem cells
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0603 Embryonic cells production of embryos, nuclear transfer A01K67/027; Embryoid bodies
• 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

Abstract

The present invention provides a preparation of undifferentiated embryonic stem (ES) cells sustainable for a prolonged period in an undifferentiated state which will undergo stem cell renewal or somatic differentiation. Preferably the cells are capable of somatic differentiation in vitro and are inclined to differentiate away from an extraembryonic lineage. The present invention also provides method of culturing embryonic stem (ES) cells to improve stem cell maintenance and persistence in culture. The method also provides a culture of ES cells prepared by the method as well as differentiated cells derived from the embryonic cells resulting from directed differentiation procedures provided by the present invention.

Claims

1. A method of preventing neuroectoderm differentiation of human embryonic stem (ES) cells, said method comprising culturing human ES cells in the presence of a noggin which is a direct antagonist of a BMP-2 mediated default pathway of extraembryonic endoderm differentiation, wherein a concentration of said noggin is in the range of 100 to 500 ng/ml to generate a population of differentiated cells that do not express Pax-6, thereby preventing neuroectoderm differentiation of human ES cells.

2. The method according to claim 1 wherein said noggin is human noggin.

3. The method according to claim 1, wherein said noggin is mouse noggin.