Development in a Primary Cell Culture of the Marine Sponge Ircinia muscarum and Analysis of the Polar Compounds

Abstract

We have established a primary cell culture of the marine demosponge Ircinia muscarum. The culture was started from a cell suspension obtained by a combination of mechanical chemical means. Microbial contamination was controlled by the use of a pool of antibiotics. Optical density, rather than hemocytometer count, is suggested to monitor the cellular growth. Analysis of the chemical composition of I. muscarum cells revealed absence of sterols, showing that the cells were unable to biosynthesize sterols. When the medium was supplemented with cholesterol an increase of about 70% in the number of cells was observed. These results suggest that the classic mammalian nutrient medium was not satisfactory for I. muscarum cell growth, and sterols were needed to satisfy the membrane requirements.

Monitoring Bacterial Diversity of the Marine Sponge Ircinia strobilina upon Transfer into Aquaculture

“…The NOMATEC (Novel Marine Technologies) project showed that Ircinia variabilis is suitable for mariculture (69). Furthermore, De Rosa et al (7) reported the development of cell cultures from Ircinia muscarum. There were major differences in the composition of secondary metabolites between the wild sponge and its cell cultures, with a lower concentration of lipids and a loss of sterols and volatile compounds in cell cultures (8).…”

Formulation of a Basal Medium for Primary Cell Culture of the Marine Sponge Hymeniacidon perleve

“…The rationale to choose these nutritional factors and the two levels for each factor (Table 2) is based on the previous studies (3)(4)(5)(6)(7)(8) as well as the composition analysis of H. perleve (16). The first screening experimental results and their statistical analysis are shown in Tables 1 and 2.…”

“…Glutamine, pyruvate, iron citrate, silicon, RPMI 1640, and Marine broth 2216 were investigated for the sponge Suberites domuncula (4). De Rosa (5,6) investigated the effects of carbon sources and cholesterols. Some growth factors tested showed positive effects on sponge cells, such as epidermal growth factor, fibroblast growth factor, insulin, wheat germ agglutinin, ulex europaeus agglutinin, phytohemagglutinin, and concanavalin A (7), but a continuous cell line is not yet established for marine invertebrates, including sponges.…”

Isolation and identification of the microalgal symbiont from primmorphs of the endemic freshwater spongeLubomirskia baicalensis(Lubomirskiidae, Porifera)

“…Half of the cultivation medium was replaced once a week. In each experiment, the following sources were used for growth: 1.0% Dulbecco's modified eagle medium (DMEM) (Sigma-Aldrich) (De Rosa et al, 2001, 2003, 1.5% fetal bovine serum (FBS) (HyClone Laboratories, Washington, U.S.A.), and different antibiotic groups: gentamycin 10 μg ml −1 (Sigma-Aldrich), a solution containing 1% penicillin (10 000 U ml −1 ) and streptomycin (10 000 μg ml −1 ) (Life Technologies, Paisley, U.K.), amphotericin 2.5 μg ml −1 (Life Technologies), and kanamycin 10 μg ml −1 (Life Technologies).…”

“…Therefore, since sponges grow comparatively slowly and sampling is often difficult, an increasing amount of attention worldwide has focused on the cultivation of sponges in vitro and the development of permanent cultures like primmorphs (Klautau et al, 1993;Ilan et al, 1996;Custodio et al, 1998;Müller et al, 1999;Rinkevich, 1999;De Rosa et al, 2001;Pomponi, 2006;Chernogor et al, 2011a). The discovery of primmorphs in freshwater sponges was made by Potts in the 19th century (Potts, 1887), when they were referred to as 'reduction bodies'.…”