Transgenic fish and a method of harvesting islet cells therefrom

• US 6,015,713 A
• Filed: 03/22/1996
• Issued: 01/18/2000
• Est. Priority Date: 04/06/1995
• Status: Expired due to Fees

• Current Assignee: Dalhousie University
• Original Assignee: Dalhousie University
• Inventors: Pohajdak, Bill, Wright, James R. Jr.
• Primary Examiner(s): Campell, Bruce R.
• Assistant Examiner(s): REYNOLDS, DEBORAH J
• Application Number:
  US08/750,391
• Time in Patent Office: 1,397 Days
• Field of Search: 424/93.1, 424/93.21, 514/44, 800/2, 800/DIG. 1-4, 530/350, 435/320.1, 435/325, 435/378
• US Class Current: 435/378
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/40 Fish
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/02 Animal zootechnically ameliorated
• CPC Class Codes: A01K 2267/025 Animal producing cells or organs for transplantation
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C07K 14/62 Insulins
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2517/02 Cells from transgenic animals

Abstract

This invention relates to transgenic fish containing a humanized insulin gene which has been altered to secrete human insulin and its use in the treatment of diabetes. In the transgenic fish of the present invention, the fish insulin gene has been modified to code for human insulin gene while leaving the regulatory sequences of fish insulin gene intact. Islet transplantation may provide the meticulous glycemia control required in the treatment of diabetes. The islet tissue of the present invention offers an inexpensive and a nearly unlimited supply of human insulin-producing tissue and therefore, may be useful in the treatment of diabetes. In this regard, an improved method of mass isolation of islet tissue is provided by the present invention. The present invention also provides the use of the humanized insulin gene to promote growth in fish.

Claims

1. A method for harvesting principal islets from fish, said method comprising:

a) incubating islet-containing tissue obtained from fish, wherein said islet-containing tissue comprises adipose tissue and connective tissue, with collagenase type II for a period of time sufficient to release the islets from adipose and connective tissue; and

b) separating the released islets from said adipose and connective tissue.

2. The method according to claim 1 wherein, in step a), the islet-containing tissue is incubated with the collagenase at 37°

C. for about 10 to about 20 minutes.