GH4C1 (ATCC® CCL-82.2)

Organism: Rattus norvegicus, rat  /  Tissue: pituitary  /  Disease: tumor

Organism Rattus norvegicus, rat
Tissue pituitary
Product Format frozen
Morphology epithelial
Culture Properties loosely adherent with floating clusters
Biosafety Level 1
Disease tumor
Age 7 months
Gender female
Strain Wistar-Furth
Applications
The GH4C1 cell line provides a system for studying the molecular mechanisms of receptor modulation and signal transduction in pituitary - derived cells.

These cells may be used for electrophysiological studies on plasma membrane calcium channels and also for studies of intracellular calcium homeostasis in secretory cells.

Cloned cDNA from GH4C1 cells has been used to investigate the structure and regulation of the rat TRH receptor.


Storage Conditions liquid nitrogen vapor phase
Karyotype modal number = 65; range = 60 to 68.
This is a hypertriploid rat cell line with the modal number of 65, occurring in 32% of cells. However, cells with 67 chromosomes also occurred at a high rate (22%). Cells with a higher ploidy count occurred at 0.6%. There were 25 to 30 marker chromosomes per cell including paired T(1;2-), single 4p+, chromosome 2 with the deleted q16-q24 (?), 11p+ and about 15 other constitutive markers. Only a single copy of the X chromosome was identified. Other characteristics included consistent single copies for chromosomes 1 and 3, double copies for chromosomes 5 and 8, and more than four copies for chromosome 7.
Derivation
The GH4C1 cell line was developed in 1972 by A.H. Tashjian, Jr. from serially propagated GH3 (ATCC CCL-82.1) cells that suddenly produced little or no detectable levels of growth hormone.

Clinical Data
female
7 months
Genes Expressed
prolactin; growth hormone (somatotrophin)
Cellular Products
prolactin; growth hormone (somatotrophin)
Comments

Note:  Cytogenetic information is based on initial seed stock at ATCC.  Cytogenetic instability has been reported in the literature for some cell lines.

GH4C1 cells differ from GH3 cells in that basal levels of growth hormone production are low (0.06 µg or less hormone/mg cell protein/24 hours).

Prolactin is also produced at a rate of 0.006 to 0.012 µg hormone/mg cell protein/24 hours.
Thyrotropin-releasing Hormone (TRH) stimulates prolactin synthesis and secretion.

These cells have a plating efficiency of approximately 22% in the recommended culture medium.
Complete Growth Medium Ham's F10 medium, 82.5%; horse serum, 15%; fetal bovine serum, 2.5%
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.. Sometimes many cells are floating, they can be harvested by centrifugation of medium instead of discarding it.
  2. Add 2.0 to 3.0 mL of 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
  3. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  4. Add appropriate aliquots of the cell suspension to new culture vessels.
  5. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:20 is recommended
Medium Renewal: 2 to 3 times per week
Cryopreservation
Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Name of Depositor AH Tashjian
References

Tashjian AH Jr., et al. Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone. Endocrinology 82: 342-352, 1968. PubMed: 4951281

Dannies PS and Tashjian AH. Growth Hormone and Prolactin from Rat Pituitary Tumor cells. Tissue Culture: methods and applications. pp. 561-569, New York: Academic Press; 1973.

Takemoto H, et al. Adrenotropic activity of mammo-somatotropic tumors in rats and mice. I. Biologic aspects. Cancer Res. 22: 917-924, 1962. PubMed: 13984648

Tashjian AH Jr.. Clonal strains of hormone-producing pituitary cells. Methods Enzymol. 58: 527-535, 1979. PubMed: 218079

Zhao D, et al. Molecular cloning of a complementary deoxyribonucleic acid encoding the thyrotropin-releasing hormone receptor and regulation of its messenger ribonucleic acid in rat GH cells [published erratum appears in Endocrinology 132: 2658, 1993]. Endocrinology 130: 3529-3536, 1992. PubMed: 1317787

Albert PR, Tashjian AH Jr.. Relationship of thyrotropin-releasing hormone-induced spike and plateau phases in cytosolic free Ca2+ concentrations to hormone secretion. Selective blockade using ionomycin and nifedipine. J. Biol. Chem. 259: 15350-15363, 1984. PubMed: 6439720

Basic Documentation
References

Tashjian AH Jr., et al. Establishment of clonal strains of rat pituitary tumor cells that secrete growth hormone. Endocrinology 82: 342-352, 1968. PubMed: 4951281

Dannies PS and Tashjian AH. Growth Hormone and Prolactin from Rat Pituitary Tumor cells. Tissue Culture: methods and applications. pp. 561-569, New York: Academic Press; 1973.

Takemoto H, et al. Adrenotropic activity of mammo-somatotropic tumors in rats and mice. I. Biologic aspects. Cancer Res. 22: 917-924, 1962. PubMed: 13984648

Tashjian AH Jr.. Clonal strains of hormone-producing pituitary cells. Methods Enzymol. 58: 527-535, 1979. PubMed: 218079

Zhao D, et al. Molecular cloning of a complementary deoxyribonucleic acid encoding the thyrotropin-releasing hormone receptor and regulation of its messenger ribonucleic acid in rat GH cells [published erratum appears in Endocrinology 132: 2658, 1993]. Endocrinology 130: 3529-3536, 1992. PubMed: 1317787

Albert PR, Tashjian AH Jr.. Relationship of thyrotropin-releasing hormone-induced spike and plateau phases in cytosolic free Ca2+ concentrations to hormone secretion. Selective blockade using ionomycin and nifedipine. J. Biol. Chem. 259: 15350-15363, 1984. PubMed: 6439720