Production technology for entomopathogenic fungus using a biphasic culture system

The objective of this research was to evaluate combinations of liquid media obtained from agro-industrial residues and by-products, with solid media prepared with mixtures of grains and their derivatives, aiming to increase the production of JAB 02 and JAB 45 isolates of Lecanicillium lecanii. Sporulation, conidial viability and process yield were evaluated as well as the production costs using the JAB 45 isolate as a model system were analyzed. The production of JAB 02 was not increased using the biphasic culture. For JAB 45, some combinations provided an increase in yield, especially cheese whey with wheat bran and wheat grain, with lower production costs. Viability was not influenced by the production method, and the combinations showed no differences in the process yield. The biphasic method is suitable for the production of L. lecanii, and proves to be an appropriate technology to use in mass production by biofactories.

Arrêté portant règlement général pour la culture du tabac en 1860. (12 décembre 1859.)

[Acte. 1859-12-12]

Arrêté portant règlement général pour la culture du tabac en 1861. (21 décembre 1860.)

[Acte. 1860-12-21]

3D culture of postnatal retinal stem cells using self assembling polymers

PURPOSE We have previously shown that retinal stem cells (RSCs) can be isolated from the radial glia population of the newborn mouse retina (Angénieux et al., 2006). These RSCs have a great capacity to renew and to generate a large number of neurons including cells differentiated towards the photoreceptor lineage (Mehri-Soussi et al., 2006). However, recent published results from our lab revealed that such cells have a poor integration and survival rate after grafting. The uncontrolled environment of a retina seems to prevent good integration and survival after grafting in vivo. To bypass this problem, we are evaluating the possibility of generating in vitro a hemi-retinal tissue before transplantation. METHODS RSC were expanded and cells passaged <10 were seeded in a solution containing poly-ethylene-glycol (PEG) polymer based hydrogels crosslinked with peptides that are chosen to be substrates for matrix metalloproteinases. Various doses of cross linkers peptides allowing connections between PEG polymers were tested. Different growth factors were studied to stimulate cell proliferation and differentiation. RESULTS Cells survived only in the presence of EGF and FGF-2 and generated colonies with a sphere shape. No cells migrated within the gel. To improve the migration and the repartition of the cells in the gels, the integrin ligand RGDSP was added into the gel. In the presence of FGF-2 and EGF, newly formed cell clusters appeared by cell proliferation within several days, but again no outspreading of cells was observed. No difference was even seen when the stiffness of the hydrogels or the concentration of the integrin ligand RGDSP were changed. However, our preliminary results show that RSCs still form spheres when laminin is entrapped in the gel, but they started to spread out having a neuronal morphology after around 2 weeks. The neuronal population was assessed by the presence of the neuronal marker b-tubulin-III. This differentiation was achieved after successive steps of stimulations including FGF-2 and EGF, and then only FGF-2. Glial cells were also present. Further characterizations are under process. CONCLUSIONS RSC can be grown in 3D. Preliminary results show that neuronal cell phenotype acquisition can be instructed by exogenous stimulations and factors linked to the gel. Further developments are necessary to form a homogenous tissue containing retinal cells.