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Primary Culture System for Germ Cells from Caenorhabditis elegans Tumorous Germline Mutants.

Bio-protocol (2017-09-05)
Alexandra S Vagasi, Mohammad M Rahman, Snehal N Chaudhari, Edward T Kipreos
RESUMO

The Caenorhabditis elegans germ line is an important model system for the study of germ stem cells. Wild-type C. elegans germ cells are syncytial and therefore cannot be isolated in in vitro cultures. In contrast, the germ cells from tumorous mutants can be fully cellularized and isolated intact from the mutant animals. Here we describe a detailed protocol for the isolation of germ cells from tumorous mutants that allows the germ cells to be maintained for extended periods in an in vitro primary culture. This protocol has been adapted from Chaudhari et al., 2016.

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Sigma-Aldrich
Tetracycline, 98.0-102.0% (HPLC)
Sigma-Aldrich
RPMI 1640 Vitamins Solution (100×), liquid, sterile-filtered, BioReagent, suitable for cell culture