Cell cultures derived from embryos and melanoma of poeciliid fish

Abstract

In poeciliid fish, melanoma of different degrees of malignancy can be produced by crossing specific genotypes. For a detailed investigation of the processes leading to proliferation or differentiation of the melanoma cells, it is necessary to establish cell cultures. The aim of the present study was to find out the optimal conditions for initiating and culturing poeciliid fish cells for the purpose of establishing cell cultures of melanoma. The optimal method was developed by using small pieces of late embryos as starting material and includes: (a) dispersion of tissue by mild stepwise treatment with a trypsin-EDTA mixture at low temperature; (b) culture of cells in the complex medium 199; (c) supplementation of medium with high percentage (20%) of fetal bovine serum; and (d) stabilization of pH by buffering the medium with HEPES. Under these conditions, primary and secondary cultures of embryonic cells have been initiated. An epithelial-like cell line has been subcultured for more than 80 passages. The method developed for embryonic tissues was used to start cell cultures from melanoma of platyfish-swordtail hybrids. Until now, only cells of rapidly growing malignant albino melanoma could be maintained in primary cultures. Secondary cultures could not be initiated since the melanoma cells tended to differentiate and stopped growing before a confluent monolayer was formed.

Gene Amplification and Functional Diversification of Melanocortin 4 Receptor at an Extremely Polymorphic Locus Controlling Sexual Maturation in the Platyfish

“…Tumor tissues were obtained from backcross hybrids between X. maculatus and X. hellerii. Xiphophorus melanoma cell line PSM (Wakamatsu 1981) and embryonic cell line A2 (Kuhn et al 1979) were used to test mc4r expression.…”

Signalling by the oncogenic receptor tyrosine kinase Xmrk leads to activation of STAT5 in Xiphophorus melanoma

“…PSM ®sh melanoma cells (Wakamatsu, 1981) expressing high amounts of endogenous Xmrk and Xiphophorus A2 embryonic cells (Kuhn et al, 1979) were cultured at 288C in F12 medium (Gibco BRL, Germany) containing 10% FCS. BHK Hm cells (Wellbrock et al, 1995) were grown in DMEM/F12 (1 : 1, Gibco BRL) with 10% FCS.…”

Activation of the Xmrk proto-oncogene of Xiphophorus by overexpression and mutational alterations

“…The cDNA of the Xmrk oncogene (Wittbrodt et al, 1989) was used to screen a cDNA library derived from the embryonal cell line A2 of X. xiphidium (Kuhn et al, 1979). Three clones were isolated which acids¯anked by 209 nucleotides of 5' untranslated sequences and 1709 nucleotides of 3' untranslated sequences.…”

“…The PSM Xiphophorus melanoma cell line (Wakamatsu, 1981) and the A2 cell line (Xiphophorus xiphidium embryonal cells, Kuhn et al, 1979) were cultured at 288C in F12 medium (Gibco BRL, Germany) containing 10% FCS. 293 cells (human embryonic kidney ®broblasts) were grown in DMEM (Gibco BRL) supplemented with 10% FCS and antibiotics.…”