Y1

Cloning and hormone studies

Mouse LAF1 adrenal cortex tumour

This cell line is a cloned strain from an adrenal cortex tumour cell line of a male LAF1 mouse. This strain was isolated after alternate passage in tumours in vivo and in cultures in vitro . The isolated clones are reported by the original depositor to produce steroid hormones at a high rate (4ug/mg/hr) under ACTH stimulation and cells round up within minutes of ACTH treatment. The hormone produced is reported to be ketosteroid with a hydroxy group at the carbon 20 rather than the usual ketosteroid configuration. However, a laboratory independent to the depositor has since (Dec 2000) tested this steroid producing property using the same method and after 1 hour of stimulation could report no rounding up or change in morphology and a very low cAMP measurement. This laboratory did find that the cells round up when exposed to forskolin (acting through adenylate cyclase) and the original depositor reported morphological changes in response to cholera toxin.

Ham′s F10 + 2mM Glutamine + 15% Horse Serum (HS) + 2.5% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:3 to 1:6 i.e. seeding at 2-4x10,000 cells/cm² using 0.25% trypsin/EDTA; 5% CO₂; 37°C.

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