Living, fast-growing thyroid cell strain, FRTL-5

• US 4,608,341 A
• Filed: 05/31/1983
• Issued: 08/26/1986
• Est. Priority Date: 05/31/1983
• Status: Expired due to Term

• Current Assignee: Interthyr Research Foundation Incorporated
• Original Assignee: Interthyr Research Foundation Incorporated
• Inventors: Ambesi-Impiombato, Francesco S.
• Primary Examiner(s): Hazel, Blondel
• Assistant Examiner(s): Tarcza, John Edward
• Application Number:
  US06/499,250
• Time in Patent Office: 1,183 Days
• Field of Search: 435/240, 435/241, 435/948, 435/244
• US Class Current: 435/353
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2501/18 Liver cell growth factor (LCGF, Gly-His-Lys)
• CPC Class Codes: C12N 2503/00 Use of cells in diagnostics
• CPC Class Codes: C12N 5/0617 Thyroid and parathyroid glands
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/78 Thyroid gland hormones , e.g. T3, T4, TBH, TBG or their receptors
• CPC Class Codes: Y10S 435/948 using viruses or cell lines

Abstract

A fast growing, continuous, functional rat thyroid cell strain, FRTL-5, which maintains functional characteristics of iodide uptake and thyroglobulin synthesis over prolonged periods of culture is cloned from FRTL cells obtained from primary cultures of Fischer rat thyroid glands. The FRTL-5 cells are cultured in a medium containing approximately 5 percent calf serum supplemented with a mixture of hormones, at least one of which is thyrotropin.

The FRTL-5 cells are employed in a series of assays which measure thyroid stimulatory or inhibitory factors. The FRTL-5 system of assay specifically measures thymidine incorporation, cAMP elevation and iodide uptake and permits the evaluation of patient sera, particularly those afflicted with Graves'"'"' disease and other autoimmune thyroid diseases, thereby providing a means for determining appropriate methods of treatment.

Claims

1. A process for preparing a continuous, functional rat cell strain, having the identifying characteristics of FRTL-5 ATCC CRL 8305 cell strain, comprising:

culturing a continuous, functional rat cell line FRTL in a medium comprising thyrotropin and calf serum to obtain FRTL-5 cells and cloning said FRTL-5 cells to obtain cells of an FRTL-5 strain having the identifying characteristics of FRTL-5 ATCC CRL 8305 cell strain.

2. The process for preparing a continuous, functional rat cell strain, FRTL-5, according to claim 1 wherein said medium is Coon'"'"'s modified Ham'"'"'s F-12 medium.

3. The process for preparing a continuous, functional rat cell strain, FRTL-5, according to claim 1 wherein said medium is supplemented with a mixture of hormones.

4. The process for preparing a continuous, functional rat cell strain, FRTL-5, according to claim 3 wherein said mixture of hormones comprises insulin, hydrocortisone, transferrin, glycyl-L-histidyl-L-lysine acetate, and somatostatin.

5. The process for preparing a continuous, functional rat cell strain, FRTL-5 according to claim 1 wherein the cell line is cultured in vitro.

6. The process for preparing a continuous, functional rat cell strain, FRTL-5, according to claim 1 wherein said calf serum is present in the range of about 2.5 to 15 percent, by volume, based on the volume of the medium.

7. The process for preparing a continuous, functional rat cell strain, FRTL-5, according to claim 6 wherein said calf serum is present in an amount of about 5 percent, by volume, based on the volume of the medium.

8. The process for preparing a continuous, functional rat cell strain FRTL-5 according to claim 1 wherein FRTL-5 cells are cloned to obtain an FRTL-5 ATCC CRL 8305 cell strain.

9. A continuous, functional rat thyroid cell strain, FRTL-5, having the accession number ATCC CRL 8305.

10. A continuous, functional rat thyroid cell strain having the identifying characteristics of FRTL-5 ATCC CRL 8305 cell strain.