Transgenic swine compositions and methods

• US 5,523,226 A
• Filed: 05/14/1993
• Issued: 06/04/1996
• Est. Priority Date: 05/14/1993
• Status: Expired due to Term

• Current Assignee: Board of Trustees of The University of Illinois
• Original Assignee: Board of Trustees of The University of Illinois, Biotechnology Research And Development Corporation (Dow, Inc.)
• Inventors: Wheeler, Matthew B.
• Primary Examiner(s): Stone, Jacqueline M.
• Assistant Examiner(s): Campell, Bruce R.
• Application Number:
  US08/063,095
• Time in Patent Office: 1,117 Days
• Field of Search: 435/6, 435/7.23, 435/172.1, 435/172.3, 435/240.2, 435/240.21, 935/70, 800/2, 800/DIG. 1
• US Class Current: 435/325
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/101 Bovine
• CPC Class Codes: A01K 2227/102 Caprine
• CPC Class Codes: A01K 2227/103 Ovine
• CPC Class Codes: A01K 2227/108 Swine
• CPC Class Codes: A01K 2267/02 Animal zootechnically ameliorated
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C12N 15/873 Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
• CPC Class Codes: C12N 2502/14 hepatocytes
• CPC Class Codes: C12N 2510/00 Genetically modified 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

Abstract

Transgenic swine, and compositions and methods for making and using same, are provided. Central to the invention are porcine (Sus scrofa) embryonic stem cell lines and methods for establishing them. Cells of such lines are transformed with exogenous genetic material of interest and then used to provide chimeric swine, which have germ cells comprising the exogenous genetic material. The chimeric swine are bred to provide transgenic swine. Transgenic swine of the invention can be used to provide human proteins or peptide hormones or can be used as xenograft donors.

Claims

1. A method of obtaining an embryonic stem cell for incorporation into a swine embryo to form a chimeric swine, said method comprising:

(a) introducing a cell from a culture made by;

(i) culturing dissociated cells from a swine embryo in conditioned stem cell medium in the presence or absence of a feeder layer, and

(ii) subculturing the culture until a stable culture with morphological features and growth parameters characteristic of an embryonic stem cell culture is established, into a SCID mouse;

(b) allowing a tumor to form in the mouse from the cell; and

(c) obtaining an embryonic stem cell from a culture that is shown to be capable of producing a tumor in step b.

2. The method of claim 1, wherein the embryonic stem cell is characterized by an undifferentiated morphology indistinguishable from the morphology of a cell from the culture of step a of claim 1 from which a cell formed a tumor in step b of claim 1.

4. An embryonic stem cell obtained from a culture that is capable of forming a tumor in a SCIDS mouse in accordance with the method of claim 1.

5. A culture initiated from an embryonic stem cell of claim 4.

3. A method for determining the cell types in which a genetic complement is expressed, said method comprising:

(a) introducing a swine embryonic stem cell Which comprises the genetic complement into an immunocompromised mouse to produce a tumor;

(b) placing the tumor in suitable conditions to allow the tumor to differentiate into a plurality of recognizable cell types and to empress the genetic complement;

(c) excising the tumor; and

(d) analyzing the differentiated cell types to determine in which cell types the genetic complement is expressed.