GT1-7 Mouse Hypothalamic GnRH Neuronal Cell Line小鼠下丘脑GnRH神经细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

Cell  Line:

GT1-7

Species:

Mus  musculus

Vulgar Name: Mouse

Tissue:

Brain

Morphology:

Fibroblast

Growth  Properties:

Adherent

Derivation:

The  GT1-7 line was immortalized by expression of SV40 T antigen oncogene in  hypothalamic GnRH neurons.

Products:

GnRH -  Gonadotropin-Releasing Hormone.

Biosafey:

2

Additional  info:

GT1-7  cell line is sensitive to trypsin and low confluence.

Culture  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, 90%; fetal bovine serum, 10%.

Subculturing:

Remove  and discard culture medium. Briefly rinse the cell layer with PBS without  calcium and magnesium 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. Add appropriate aliquots of the cell suspension to new culture  vessels. Incubate cultures at 37°C. Doubling time: 36 hours NOTE: For more  information on enzymatic dissociation and subculturing of cell lines consult  Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian  Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.

Medium Renewal: Every 2 to 3 days

Subcultivation ratio: 1:5 to 1:7

Culture  Conditions:

Atmosphere:  air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C

Cryopreservation:

95% FBS  + 5% DMSO (Dimethyl sulfoxide)

Thawing  Frozen Cells:

SAFETY  PRECAUTION: Is highly recommend that protective gloves and clothing always be  used and a full face mask always be worn when handling frozen vials. It is  important to note that some vials leak when submersed in liquid nitrogen and  will slowly fill with liquid nitrogen. Upon thawing, the conversion of the  liquid nitrogen back to its gas phase may result in the vessel exploding or  blowing off its cap with dangerous force creating flying debris. 1. Thaw the  vial by gentle agitation in a 37°C water bath. To reduce the possibility of  contamination, keep the Oring and cap out of the water. Thawing should be rapid  (approximately 2 minutes). 2. Remove the vial from the water bath as soon as  the contents are thawed, and decontaminate by dipping in or spraying with 70%  ethanol. All of the operations from this point on should be carried out under  strict aseptic conditions. 3. For cells that are sensitive to DMSO is  recommended that the cryoprotective agent be removed immediately. Transfer  the vial contents to a centrifuge tube containing 9.0 mL complete culture  medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the  supernatant and Resuspend cell pellet with the recommended complete medium  (see the specific batch information for the culture recommended dilution  ratio). 5. Incubate the culture in a appropriate atmosphere and temperature  (see "Culture Conditions" for this cell line). NOTE: It is  important to avoid excessive alkalinity of the medium during recovery of the  cells. It is suggested that, prior to the addition of the vial contents, the  culture vessel containing the growth medium be placed into the incubator for  at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

References:

Nelson  SB, LAwson MA, Kelley CG, Mellon PL (2000) "neuron-specific expression  of the rat gonadotropin-releasing hormone gene is confered by interactions of  a defined promoter with the enhacer in GT1-7 cells". Mol Endocrinol 14  (9): 1509-22. Mellon, P.L., Windle, J,J., Goldsmith, P., Pedula, C., Robets,  J. and Weiner, R.I. 1990. Imortalization of hypothalamic GnRH neurons by  genetically targeted tumorigenesis. Neuron 5: 1-10. Liposits, Z.,  Mechenthaler,I., Westel, W.C., Reid, J. J., Mellon, P. L., Weiner, R.I. and  Negro-Villar, A.1991. Morphological chaacterization of immortalized  hypothalamic neurons synthesizing luteinizing hormone-releasing hormone.  Endocrinol. 129: 1575-1583. Wetsel, W.C., Valença, M.M., Merchenthaler, I.  Liposits, Z., López, F.J., Weiner, R.I., Mellon, P.L. and Negro-Villar, A.  1992. Intrinsic pulsatile secretory activity of immortalized luteinizing  hormone-releasing hormone-secreting neurons. Proc. Natl. Acad. Sci. USA  89:4149-4153. Whyte, D. B., Lawson, M.A., Belsham,D. D., Eraly, S.A. Bond,  C.T., Adelman, J. P., and Mellon, P.L. 1995. A Neuron-Specific Enhancer  Targets Expression of the Gonadotropin-Releasing Hormone Gene to Hypothalamic  Neurosecretory Neurons. Molecular Endocrinology 9, 467-477.