Meeting Abstract
Primary cell cultures are derived from tissue explants removed from an organism and maintained in an artificial environment. These in vitro cell cultures can be used to approximate the in vivo cellular environment, facilitate the isolation of specific cell types, and often provide a better representation of normal cell biology than immortalized cell lines. We have developed a primary cell culture system for the marine invertebrate Mnemiopsis leidyi, a lobate ctenophore. These primary cell cultures can serve as a proxy for adult somatic cells. Treated media derived from adult animals is used to generate both a solid matrix for cell adhesion and support as well as a liquid overlay. This complex media provides essential nutrients to isolated explant cells. Approximately 24 hours after explant removal several different cell morphologies are consistently observed. Many cells have cytoplasmic processes and are highly motile. Extensive, active cell processes often connect cells. A distinct class of round proliferative cells characterized by a lack of cytoplasmic processes are also reliably isolated. We are currently employing several methods to further characterize these cells that can be loosely classified by morphology as either round, bipolar, multipolar, or hyper-elongated cells. A rt-PCR approach is being used to develop gene expression profiles associated with distinct cell morphologies as well as changes in gene expression over time. We are also performing experiments to monitor the functional response of Calcium channels via trafficking of labeled small molecules. Experiments to further characterize discrete cell types associated with these primary cell cultures will facilitate our ongoing molecular genetic analysis of unique aspects of ctenophore cell differentiation during development and ctenophore evolutionary history from a cell biological perspective.
