KG-1a

After the tenth passage, the parental KG-1 cells were cultured in two separate laboratories within the same department under identical conditions.

After 35 passages the cells in one laboratory expressed morphological differences from the parent line. The variant KG-1a was composed of undifferentiated promyeloblasts.

Both populations exhibit many common characteristics. They share a similar doubling time, are negative for EBNA and VCA, express no surface immunoglobulins and exhibit identical HLA and isoenzyme profiles.

In contrast to the parental KG-1 (ATCC CCL-246) the KG-1a population is unresponsive to colony-stimulating factor in soft-agar culture and does not express the Ia-like antigen.

KG-1a cells are resistant to phorbol diester induced macrophage differentiation and proliferation of the cells is unaffected by the presence of phorbol diesters

.The KG-1a cells are morphologically, cytochemically, and functionally less mature than the parental KG-1.

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.

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

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.        

Maintain cell density between 2 x 10⁵ and 1 x 10⁶ viable cells/mL.

Medium Renewal: Twice per week

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