RC-4B/C細(xì)胞

運輸方式： 凍存運輸

ATCC Number： CRL-1903?

相關(guān)**： 其他**

生長狀態(tài)： 貼壁生長

數(shù)量： 大量

細(xì)胞形態(tài)： 上皮樣

器官來源： 其他

年限： 3 years

是否是腫瘤細(xì)胞： 0

物種來源： 大鼠

RC-4B/C細(xì)胞Designations: RC-4B/C

Depositors: EH Leiter

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Rattus norvegicus deposited as Rattus sp.

Morphology: epithelial

Source: Organ: pituitary, anterior

Disease: adenoma

Cellular Products: luteinizing hormone beta (LH); growth hormone (GH); follicle stimulating hormone beta (FSH); prolactin; adrenal corticotropic hormone (ACTH); thyrotropin beta

Permits/Forms: RC-4B/C細(xì)胞In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

Receptors: gonadotropin releasing hormone (GnRH)

Age: 3 years

Gender: male

Comments: This line was carried continuously in culture for 5 years before it was sucesssfully cryopreserved.

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.

Propagation: RC-4B/C細(xì)胞ATCC 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%

Temperature: 37.0°C

Subculturing: Protocol:

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.

RC-4B/C細(xì)胞To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg 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

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

References: 22681: 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