Hormone-secreting cells derived from pancreatic islet maintained in long-term culture

• US 5,928,942 A
• Filed: 06/07/1995
• Issued: 07/27/1999
• Est. Priority Date: 06/24/1991
• Status: Expired due to Fees

• Current Assignee: Pacific Biomedical Research Incorporated
• Original Assignee: Pacific Biomedical Research Incorporated
• Inventors: Brothers, Ann Janice
• Primary Examiner(s): Saucier, Sandra E.
• Application Number:
  US08/475,969
• Time in Patent Office: 1,511 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/347, 435/371, 435/373, 435/377, 435/70.3
• US Class Current: 435/347
• 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: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/33 other than alpha-amino carboxylic acids, e.g. beta-amino acids, taurine
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/31 Pituitary sex hormones, e.g. follicle-stimulating hormone [FSH], luteinising hormone [LH]; Chorionic gonadotropins
• CPC Class Codes: C12N 2503/00 Use of cells in diagnostics
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0617 Thyroid and parathyroid glands
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0682 Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells oocytes C12N5/0609
• CPC Class Codes: C12P 21/02 having a known sequence of two or more amino acids, e.g. glutathione
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025

Abstract

Methods are provided for the establishment and maintenance in long term culture of hormone secreting cells. Cells are derived from tumorous or non-tumorous animal or human tissues, including ovary, endometrium, trophoblast, pituitary, thyroid, and pancreas. The cells secrete into the culture medium hormones such as estrogens, progestins, follicle-stimulating hormone, luteinizing hormone, human chorionic gonadotrophin, thyroxin, glucagon, and insulin, depending on the tissue of origin of individual cell cultures. Contact with an appropriate secretogogue causes the cells to respond with increased hormone secretion. For instance, ovarian follicular cells respond to follicle-stimulating hormone with increased estrogen and progesterone secretion. Pancreatic cells respond to elevated glucose with increased insulin secretion. The cells proliferate in in vitro for up to one year or longer, during which time they retain their hormone-secretion profile. The cells may be frozen for storage, and retain their hormone-secretion profile after thawing. The cell cultures are useful for the production of human hormones, for the bio-assay of drugs such as therapeutic gonadotrophins, for the testing of drug efficacy and design, and for toxicity testing of drugs and chemicals. The cells may also be implanted in an individual to replace deficient hormone secretion. For instance, insulin secreting pancreatic cells may be implanted in a diabetic individual as an adjunct or replacement therapy for exogenously administered insulin.

Claims

1. A cell culture of replicating, hormone-secreting cells wherein the cell culture has been continuously maintained for at least about three months, said cells are derived from at least one isolated, normal human pancreatic islet cell, which has not been subjected to treatment with trypsin and wherein said isolated pancreatic islet cell existed in a nodule formed from the protrusion of islet cells through a deliberately manually, mechanically or enzymatically caused rupture in the basement lamina of the isolated human pancreatic whole islet, wherein said cultured cells secrete insulin in the amount of about 2 to about 150 μ

IU per milliliter per hour per thousand cells in a glucose dose-related response when said cells are contacted with 6 to about 22 mM glucose.

2. The culture according to claim 1 wherein said culture has been continuously maintained for at least nine months.

3. The culture according to claim 1 wherein said cells proliferate at a rate such that there is about a 20-fold increase in the cell number after about three to about six months of culture.

4. The culture according to claim 1 wherein said cells proliferate at a rate such that about 30% to about 90% of said culture cells enter the mitotic cycle about every 10 days.

5. The culture according to claim 1 wherein said culture has been maintained for at least 17 months.

6. The culture according to claim 1 wherein said culture has been maintained for at least 12 months.