Claims
- 1. A method for establishing, maintaining and propagating a cell culture of gonadotrophin-secreting human pituitary tumor cells in vitro comprising the steps of:a) selecting at least one human pituitary tumor cell having gonadotrophin-secreting potential from a population of similar cells having gonadotrophin-secreting potential, b) placing said cell or cells in an establishing medium comprising: (i) a basal medium comprising essential minerals, salts, vitamins, amino acids, and lipids; (ii) a buffering system; (iii) glutamine in the amount of about 6.18 mM to about 8.36 mM; (iv) pyruvate; (v) niacinamide; (vi) myo-Inositol; and (vii) supplements of a fibroblast growth factor, an epidermal growth factor, insulin, ethanolamine, selenium, transferrin and hydrocortisone, said establishing medium promoting the viability and proliferation of said cell or cells for at least about 13 days in vitro, and c) maintaining the viability of at least one of said cells or the progeny of said cell for at least about 15 days in vitro wherein said viable cell secretes or viable cells secrete at least 4.04 mIU of follicle stimulating hormone (FSH)/ml establishing medium or at least 11.3 mIU of luteinizing hormone (LH)/ml establishing medium over a 10-day synthesis time in a substantially estrogen free culture.
- 2. A method for establishing, maintaining and propagating a cell culture of gonadotrophin-secreting human pituitary tumor cells in vitro comprising the steps of:a) selecting at least one human pituitary tumor cell having gonadotrophin-secreting potential from a population of similar cells having gonadotrophin-secreting potential, b) placing said cell or cells in an establishing medium comprising: (i) a basal medium comprising essential minerals, salts, vitamins, amino acids, and lipids; (ii) a buffering system; (iii) glutamine in the amount of about 6.18 mM to about 8.36 mM; (iv) pyruvate; (v) niacinamide; (vi) myo-Inositol; and (vii) supplements of a fibroblast growth factor, an epidermal growth factor, insulin, ethanolamine, selenium, transferrin and hydrocortisone, said establishing medium promoting the viability and proliferation of said cell or cells for at least about 13 days in vitro, and c) maintaining the viability of at least one of said cells or the progeny of said cell for at least about 15 days in vitro wherein said viable cell secretes or viable cells secrete at least 4.04 mIU FSH/ml establishing medium and at least 11.3 mIU LH/ml establishing medium over a 10 day synthesis time in a substantially estrogen free culture.
- 3. A method for establishing, maintaining and propagating a cell culture of gonadotrophin-secreting human pituitary tumor cells in vitro comprising the steps of:a) selecting at least one human pituitary tumor cell having gonadotrophin-secreting potential from a population of similar cells having gonadotrophin-secreting potential, b) placing said cell or cells in an establishing medium comprising: (i) a basal medium comprising essential minerals, salts, vitamins, amino acids, and lipids; (ii) a buffering system; (iii) glutamine in the amount of about 6.18 mM to about 8.36 m; (iv) pyruvate; (v) niacinamide; (vi) myo-Inositol; and (vii) supplements of a fibroblast growth factor, an epidermal growth factor, insulin, ethanolamine, selenium, transferrin and hydrocortisone, said establishing medium promoting the viability and proliferation of said cell or cells for at least about 13 days in vitro, and c) maintaining the viability of at least one of said cells or the progeny of said cell for 28 days in vitro wherein said viable cell secretes or viable cells secrete 3.0 mIU LH/ml establishing medium in a substantially estrogen free culture over 8 days.
CROSS-REFERENCE TO RELATED APPLICATION
This application is a continuation of U.S. patent application Ser. No. 08/475,969 filed Jun. 7, 1995, now U.S. Pat. No. 5,928,942, issued Jul. 27, 1999, which is a continuation-in-part of U.S. patent application Ser. No. 08/167,862, filed Apr. 29, 1994 (now abandoned), which is a 371 national stage entry of PCT/US92/05267, filed Jun. 23, 1992, entitled “Hormone-Secreting Cells Maintained in Long-Term Cultures” which is a continuation-in-part of U.S. patent application Ser. No. 07/719,977, filed Jun. 24, 1991, now abandoned, entitled “Hormone Secreting Cells Maintained in Long-Term Culture for the Production of Hormones, for Bio-Assay of Drugs, and for Toxicity Testing of Drugs and Chemicals”. Related applications are Ser. No. 08/477,067, filed Jun. 7, 1995, now U.S. Pat. No. 5,821,121, issued Oct. 13, 1998, and Ser. No. 08/486,737, filed Jun. 7, 1995, now U.S. Pat. No. 5,787,341, issued May 5, 1998.
US Referenced Citations (2)
Number |
Name |
Date |
Kind |
3882233 |
Grant et al. |
May 1975 |
A |
4288546 |
Narasimhan et al. |
Sep 1981 |
A |
Non-Patent Literature Citations (2)
Entry |
Goodyer et al. J. Clin. Endocrinol. Metabolism. vol. 45, No. 1, pp. 73-85, 1977.* |
Lipson et al. Acta Endocrinol. vol. 90, No. 3, pp. 421-433, 1979. |
Continuations (1)
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08/475969 |
Jun 1995 |
US |
Child |
09/301110 |
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US |
Continuation in Parts (2)
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Date |
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08/167862 |
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US |
Child |
08/475969 |
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US |
Parent |
07/719977 |
Jun 1991 |
US |
Child |
08/167862 |
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US |