Establishment of a human embryonic stem cell line using mammalian cells

• US 7,811,817 B2
• Filed: 05/17/2006
• Issued: 10/12/2010
• Est. Priority Date: 05/17/2005
• Status: Expired due to Fees

• Current Assignee: Reliance Life Sciences Pvt. Ltd.
• Original Assignee: Reliance Life Sciences Pvt. Ltd.
• Inventors: Ravindran, Geeta, Kulkarni, Jayant, Bose, Bipasha, Patki, Ameet, Kahn, Alam Firdos, Pal, Rajarshi, Mandal, Arundhati, Khanna, Aparna, Tipnis, Shabari
• Primary Examiner(s): Nguyen; Quang
• Application Number:
  US11/436,193
• Publication Number:
  US 20060286544A1
• Time in Patent Office: 1,609 Days
• Field of Search: None
• US Class Current: 435/366
• 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

Purified preparations of human embryonic stem cells with certain population-specific characteristics are disclosed. This preparation is characterized by the positive expression of the following pluripotent cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); alkaline phosphatase (+), as well as a set of ES cell markers including Oct-4, Nanog, Rex1, Sox2, Thy1, FGF4, ABCG2, Dppa5, UTF1, Cripto1, hTERT, Connexin-43 and Connexin-45. The cells of the preparation are negative for lineage specific markers like Keratin 8, Sox-1, NFH (ectoderm), MyoD, brachyury, cardiac-actin (mesoderm), HNF-3 beta, albumin, and PDX1 (endoderm). The cells of the preparation are human embryonic stem cells, have normal karyotypes, exhibit high telomerase activity and continue to proliferate in an undifferentiated state after continuous culture for over 40 passages. The embryonic stem cell line Relicell™ hES1 also retains the ability, throughout the culture, to differentiate into cell and tissue types derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). Methods for isolating a human embryonic stem cell line are also disclosed.

Claims

1. An isolated pluripotent human embryonic stem cell line deposited as ATCC Patent Deposit Designation No. PTA-8172.

2. A method of screening a substance for its effect on a purified preparation of pluripotent human embryonic stem cells, comprising:

a) obtaining the purified preparation of pluripotent human embryonic stem cells;

b) combining the preparation of pluripotent human embryonic stem cells with the substance; and

c) determining any effect of the substance on the cells in the preparation;

wherein the purified preparation of pluripotent human embryonic stem cells is from the pluripotent human embryonic stem cell line deposited as ATCC Patent Deposit Designation No. PTA-8172.