Method of producing an avian embryonic stem cell culture and the avian embryonic stem cell culture produced by the process

• US 5,340,740 A
• Filed: 05/15/1992
• Issued: 08/23/1994
• Est. Priority Date: 05/15/1992
• Status: Expired due to Term

• Current Assignee: North Carolina State University (University of North Carolina System)
• Original Assignee: North Carolina State University (University of North Carolina System)
• Inventors: Petitte, James N., Yang, Zengming
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): DADIO, SUSAN
• Application Number:
  US07/884,423
• Time in Patent Office: 830 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/240.3, 435/240.31, 424/93 U
• US Class Current: 435/349
• CPC Class Codes: A01K 2227/30 Bird
• CPC Class Codes: A61K 35/57 Birds; Materials from birds, e.g. eggs, feathers, egg white, egg yolk or endothelium corneum gigeriae galli
• CPC Class Codes: C12N 2502/13 connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
• CPC Class Codes: C12N 2502/14 hepatocytes
• 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

A method of producing undifferentiated avian cells expressing an embryonic stem cell phenotype is disclosed. The method comprises collecting avian cells from an avian blastoderm prior to formation of the primitive streak, then depositing the avian cells in contact with a mouse fibroblast feeder cell layer, and then growing the avian cells on the mouse fibroblast feeder cell layer in the presence of a media containing leukemia inhibitory factor in a differentiation-inhibiting amount for a time sufficient to produce a sustained avian cell culture. Cell cultures produced by the aforesaid process and veterinary pharmaceutical formulations containing such cells are also disclosed.

Claims

1. A method of producing a sustained culture of undifferentiated avian cells expressing an embryonic stem cell phenotype, comprising:

collecting avian cells from an avian blastoderm prior to formation of the primitive streak;

depositing said avian cells in contact with a mouse fibroblast feeder cell layer;

growing said avian cells on said mouse fibroblast feeder cell layer in the presence of a media containing leukemia inhibitory factor in a differentiation-inhibiting amount for a time sufficient to produce a sustained avian cell culture;

said sustained avian cell culture consisting essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype.

2. A method according to claim 1, wherein said avian cells are collected from the area pellucida of a stage IX to stage XIV blastoderm.

3. A method according to claim 1, wherein said undifferentiated avian cells are capable of maintaining said stem cell phenotype when grown on said mouse fibroblast feeder layer in the presence of said media for at least three days.

4. A method according to claim 1, wherein said avian blastoderm is a blastoderm selected from the group consisting of chicken, turkey, duck, geese, quail and pheasant blastoderms.

5. A method according to claim 1, wherein said avian blastoderm is a chicken blastoderm.

6. A method according to claim 1, wherein said media is a conditioned media.

7. A method according to claim 1, wherein said mouse fibroblast feeder cell layer is a mouse STO fibroblast feeder cell layer.

8. A sustained avian cell culture of undifferentiated avian cells expressing an embryonic stem cell phenotype produced by the process of claim 1.

9. A method of producing a sustained culture of undifferentiated chicken cells expressing an embryonic stem cell phenotype, comprising:

collecting avian cells from the area pellucida of a stage IX to stage XIV chicken blastoderm;

depositing said avian cells on contact with a mouse STO fibroblast feed cell layer;

growing said avian cells on said mouse fibroblast feeder cell layer in the presence of a conditioned media containing leukemia inhibitory factor in a differentiation-inhibiting amount of a time sufficient to produce a sustained avian cell culture;

said sustained avian cell culture consisting essentially of undifferentiated avian cells expressing an embryonic stem cell phenotype, and wherein said undifferentiated avian cells are capable of maintaining said stem cell phenotype when grown on said mouse fibroblast feeder layer in the presence of said media for at least three days.

10. A sustained chicken cell culture of undifferentiated chicken cells expressing an embryonic stem cell phenotype produced by the process of claim 9.