Human embryonic stem cells and culturing methods thereof

• US 7,682,826 B2
• Filed: 09/23/2005
• Issued: 03/23/2010
• Est. Priority Date: 12/09/2003
• Status: Expired due to Fees

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Yang, Mei Ju, Chen, Wann Hsin, Huang, Jun Jae, Kuo, Chao Ying, Hsu, Lih Tao, Chang, Kuang Ning
• Primary Examiner(s): DRISCOLL, LORA E BARNHART
• Application Number:
  US11/233,055
• Publication Number:
  US 20060030040A1
• Time in Patent Office: 1,642 Days
• Field of Search: None
• US Class Current: 435/377
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• 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

Provided herein are human embryonic stem cells and their method of culturing. In one embodiment, the human embryonic stem cells are maintained in an environment containing an extracellular matrix isolated from feeder cells and a conditioned medium pre-conditioned by the feeder cells; the feeder cells are pre-inactivated by either gamma ray radiation or by mitomycin C treatment. The cultured human embryonic stem cells remain substantially undifferentiated and maintain their pluripotency to differentiate into three germ layer cells.

Claims

1. A composition for maintaining the undifferentiated growth of human embryonic stem cells, the composition comprising:

a substrate covered with an extracellular matrix isolated from inactivated feeder cells;

a conditioned medium having been preconditioned by inactivated feeder cells; and

undifferentiated human embryonic stem cells;

wherein the feeder cells are selected from the group consisting of human foreskin fibroblasts and primary mouse embryonic fibroblasts and

wherein the feeder cells have been inactivated by treatment with 10μ

g/mL mitomycin C.