KC-4031

Ba/F3-rat-GHR-High Cell Line

40520

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Background of Ba/F3-rat-GHR-High Cell Line

GHR is growth hormone receptor, which induces differentiation mature, starts the synthesis in the cell metabolism, and promotes cell proliferation. GHR is associated with a variety of tumors. GHR knockout promotes apoptosis of gastric cancer cells. GHR regulates G1 phase cell cycle progression by regulating the PI3K/AKT signaling pathway, thereby regulating the growth and apoptosis of gastric cancer cells. The findings in the treatment of gastric cancer provides a promising new strategy. Diseases associated with GHR include Laron Syndrome and Growth Hormone Insensitivity, Partial.

Specifications

Catalog Number	KC-4031
Cell Line Name	Ba/F3-rat-GHR-High Cell Line
Clone Number	8#
Host Cell Line	Ba/F3
Description	Stable Ba/F3 clone expressing exogenous rat GHR gene in high level
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640 + 10% FBS+1μg/mL puromycin+8ng/mL IL-3
Selection Marker	Puromycin
Morphology	Mostly single, round (some polymorph) cells in suspension
Subculture	Split saturated culture 1:10 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 20 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Ba/F3-rat-GHR-High cell line was generated using a lentiviral vector expressing the rat GHR sequence.

Characterization

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 + 10% FBS+1μg/mL puromycin+8ng/mL IL-3)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:10 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

References

1. Yan HZ, Wang HF, Yin Y, Zou J, Xiao F, Yi LN, He Y, He BS. GHR is involved in gastric cell growth and apoptosis via PI3K/AKT signalling. J Cell Mol Med. 2021 Mar;25(5):2450-2458. doi: 10.1111/jcmm.16160. Epub 2021 Jan 25. PMID: 33492754; PMCID: PMC7933969. 2. Guevara-Aguirre J, Guevara A, Palacios I, Pérez M, Prócel P, Terán E. GH and GHR signaling in human disease. Growth Horm IGF Res. 2018 Feb;38:34-38. doi: 10.1016/j.ghir.2017.12.006. Epub 2017 Dec 12. PMID: 29395968. 3. de Lima JBM, Debarba LK, Rupp AC, Qi N, Ubah C, Khan M, Didyuk O, Ayyar I, Koch M, Sandoval DA, Sadagurski M. ARCGHR Neurons Regulate Muscle Glucose Uptake. Cells. 2021 May 3;10(5):1093. doi: 10.3390/cells10051093. PMID: 34063647; PMCID: PMC8147615.