GH3

Although the GH3 was not a direct derivative of the GH1 clone (ATCC CCL-82), it was initiated after only two additional animal passages from the primary culture from which the GH1 clone was established.

The cell line was submitted to the Ameri

The epithelial-like GH3 clone generates growth hormone at a greater rate than the GH1 cells and also produces prolactin.

Studies on the control of the production of these protein hormones by the GH3 cells have shown that hydrocortisone stimulates the production of growth hormone and inhibits prolactin production.

The cells have been adapted to growth in suspension culture using Eagle's minimum essential medium (spinner) supplemented with 15% horse serum and 2.5% FBS. Under these conditions the cells continue to produce both growth hormone and prolactin

1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.

1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring 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. 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. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 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 complete 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). pH (7.0 to 7.6).
5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO₂ in air atmosphere is recommended if using the medium described on this product sheet.

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.

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