| 產(chǎn)品名稱 |
RC-4B/C |
| 商品貨號(hào) |
B165585 |
| Organism |
Rattus norvegicus, rat |
| Tissue |
pituitary, anterior |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
adenoma |
| Age |
3 years |
| Gender |
male |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
This line was carried continuously in culture for 5 years before it was sucesssfully cryopreserved. |
| Clinical Data |
male |
| Receptor Expression |
gonadotropin releasing hormone (GnRH) |
| Genes Expressed |
luteinizing hormone beta (LH); growth hormone (GH); follicle stimulating hormone beta (FSH); prolactin; adrenal corticotropic hormone (ACTH); thyrotropin beta |
| Cellular Products |
luteinizing hormone beta (LH); growth hormone (GH); follicle stimulating hormone beta (FSH); prolactin; adrenal corticotropic hormone (ACTH); thyrotropin beta |
| Comments |
The line contains cells producing LH, GH, FSH, prolactin, ACTH and thyrotropin beta (hormone production was tested using cells that were grown in medium containing Nu-serum rather than fetal bovine serum).
GnRH receptors are about 2 fold lower in number than on normal pituitary.
The cells contain numerous electron dense secretory granules, golgi complexes, and extended arrays of rough endoplasmic reticulum as well as endogenous rat C-type retrovirus particles.
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| Complete Growth Medium |
Dulbecco's modified Eagle's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate and 4.5 g/L glucose, 45%; alpha minimum essential medium with 1 g/L glucose, 45%; supplemented with 0.01 mM nonessential amino acids, 15 mM HEPES, 0.2 mg/ml bovine serum albumin, 2.5 ng/ml epidermal growth factor; dialyzed heat-inactivated fetal bovine serum, 10%
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| 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.
- Remove and discard culture medium.
-
Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
-
Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
-
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
-
To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
EH Leiter |
| Deposited As |
Rattus sp. |
| References |
Hurbain-Kosmath I, et al. Gonadotropes in a novel rat pituitary tumor cell line, RC-4B/C. Establishment and partial characterization of the cell line. In Vitro Cell. Dev. Biol. 26: 431-440, 1990. PubMed: 2161825
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