Methods and compositions for feeder-free pluripotent stem cell media containing human serum

• US 9,267,110 B2
• Filed: 12/10/2013
• Issued: 02/23/2016
• Est. Priority Date: 10/19/2007
• Status: Active Grant

• Current Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Viacyte Incorporated (Vertex Pharmaceuticals Incorporated)
• Inventors: Robins, Allan J, Schulz, Thomas C
• Primary Examiner(s): Ton, Thaian N
• Assistant Examiner(s): Moloye, Titilayo
• Application Number:
  US14/102,451
• Publication Number:
  US 20140099713A1
• Time in Patent Office: 805 Days
• US Class Current: 1/1
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2500/98 Xeno-free medium and culture conditions
• CPC Class Codes: C12N 2501/15 Transforming growth factor beta (TGF-β)
• 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 5/068 Stem cells; Progenitors

Abstract

The present invention provides compositions and methods for the culture and maintenance of pluripotent stem cells. More particularly, the present invention provides for compositions and methods for culturing, maintaining, growing and stabilizing primate pluripotent stem cells in a feeder-free defined media further comprising human serum, or a soluble attachment component of the human serum, for promoting cell attachment.

Claims

1. A composition comprising primate pluripotent cells, and a medium comprising an effective amount of human serum in a matrix-free culture vessel, wherein the primate pluripotent cells are in contact with the medium in the culture vessel and wherein the serum promotes attachment of the primate pluripotent cells and the cells are attached to the culture vessel.

2. The composition of claim 1, wherein the medium supports proliferation of the primate pluripotent cell in an undifferentiated state.

3. The composition of claim 1, wherein the culture vessel does not comprise feeder cells.

4. The composition of claim 1, wherein the culture vessel is a bioreactor.

5. The composition of claim 1, wherein the primate pluripotent cells are human embryonic stem cells (hESC).

6. The composition of claim 1, wherein the human serum is at a concentration of about 0.1% to about 20% of the medium.

7. The composition of claim 1, wherein the primate pluripotent stem cells continue to proliferate without differentiation for more than 2 serial passages.

8. The composition of claim 1, wherein the primate pluripotent stem cells are passaged by enzymatic dissociation and replating.