Human pluripotent stem cells (hPSCs) have gained a solid foothold in basic research and drug industry as they can be used in vitro to study human development and have potential to offer limitless supply of various somatic cell types needed in drug development. Although the hepatic differentiation of hPSCs has been extensively studied, only a little attention has been paid to the role of the extracellular matrix. In this study we used laminin-511, laminin-521, and fibronectin, found in human liver progenitor cells, as culture matrices for hPSC-derived definitive endoderm cells. We observed that laminin-511 and laminin-521 either alone or in combination support the hepatic specification and that fibronectin is not a vital matrix protein for the hPSC-derived definitive endoderm cells. The expression of the laminin-511/521-specific integrins increased during the definitive endoderm induction and hepatic specification. The hepatic cells differentiated on laminin matrices showed the upregulation of liver-specific markers both at mRNA and protein levels, secreted human albumin, stored glycogen, and exhibited cytochrome P450 enzyme activity and inducibility. Altogether, we found that laminin-511 and laminin-521 can be used as stage-specific matrices to guide the hepatic specification of hPSC-derived definitive endoderm cells.
- MeSH
- biomimetické materiály chemie MeSH
- buněčná diferenciace fyziologie MeSH
- buněčné linie MeSH
- extracelulární matrix - proteiny metabolismus MeSH
- extracelulární matrix metabolismus MeSH
- hepatocyty cytologie fyziologie MeSH
- laminin metabolismus MeSH
- lidé MeSH
- pluripotentní kmenové buňky cytologie metabolismus MeSH
- techniky vsádkové kultivace metody MeSH
- tkáňové inženýrství metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Penicillin production during a fermentation process using industrial strains of Penicillium chrysogenum is a research topic permanently discussed since the accidental discovery of the antibiotic. Intact cell mass spectrometry (ICMS) can be a fast and novel monitoring tool for the fermentation progress during penicillin V production in a nearly real-time fashion. This method is already used for the characterization of microorganisms and the differentiation of fungal strains; therefore, the application of ICMS to samples directly harvested from a fermenter is a promising possibility to get fast information about the progress of fungal growth. After the optimization of the ICMS method to penicillin V fermentation broth samples, the obtained ICMS data were evaluated by hierarchical cluster analysis or an in-house software solution written especially for ICMS data comparison. Growth stages of a batch and fed-batch fermentation of Penicillium chrysogenum are differentiated by one of those statistical approaches. The application of two matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) instruments in the linear positive ion mode from different vendors demonstrated the universal applicability of the developed ICMS method. The base for a fast and easy-to-use method for monitoring the fermentation progress of P. chrysogenum is created with this ICMS method developed especially for fermentation broth samples.
Clostridium acetobutylicum immobilised in polyvinylalcohol, lens-shaped hydrogel capsules (LentiKats(®)) was studied for production of butanol and other products of acetone-butanol-ethanol fermentation. After optimising the immobilisation protocol for anaerobic bacteria, continuous, repeated batch, and fed-batch fermentations in repeated batch mode were performed. Using glucose as a substrate, butanol productivity of 0.41 g/L/h and solvent productivity of 0.63 g/L/h were observed at a dilution rate of 0.05 h(-1) during continuous fermentation with a concentrated substrate (60 g/L). Through the process of repeated batch fermentation, the duration of fermentation was reduced from 27.8h (free-cell fermentation) to 3.3h (immobilised cells) with a solvent productivity of 0.77 g/L/h (butanol 0.57 g/L/h). The highest butanol and solvent productivities of 1.21 and 1.91 g/L/h were observed during fed-batch fermentation operated in repeated batch mode with yields of butanol (0.15 g/g) and solvents (0.24 g/g), respectively, produced per gram of glucose.
- MeSH
- aceton metabolismus MeSH
- anaerobióza MeSH
- butanoly metabolismus MeSH
- Clostridium acetobutylicum cytologie metabolismus MeSH
- ethanol metabolismus MeSH
- fermentace * MeSH
- imobilizované buňky metabolismus MeSH
- techniky vsádkové kultivace metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of this work was to study the effect of Se(+VI) on viability, cell morphology, and selenomethionine accumulation of the green alga Chlorella sorokiniana grown in batch cultures. Culture exposed to sublethal Se concentrations of 40 mg · L(-1) (212 μM) decreased growth rates for about 25% compared to control. A selenate EC50 value of 45 mg · L(-1) (238.2 μM) was determined. Results showed that chlorophyll and carotenoids contents were not affected by Se exposure, while oxygen evolution decreased by half. Ultrastructural studies revealed granular stroma, fingerprint-like appearance of thylakoids which did not compromise cell activity. Unlike control cultures, SDS PAGE electrophoresis of crude extracts from selenate-exposed cell cultures revealed appearance of a protein band identified as 53 kDa Rubisco large subunit of Chlorella sorokiniana, suggesting that selenate affects expression of the corresponding chloroplast gene as this subunit is encoded in the chloroplast DNA. Results revealed that the microalga was able to accumulate up to 140 mg · kg(-1) of SeMet in 120 h of cultivation. This paper shows that Chlorella sorokiniana biomass can be enriched in the high value aminoacid SeMet in batch cultures, while keeping photochemical viability and carbon dioxide fixation activity intact, if exposed to suitable sublethal concentrations of Se.
- MeSH
- bioreaktory mikrobiologie MeSH
- Chlorella cytologie účinky léků fyziologie MeSH
- kyselina selenová aplikace a dávkování MeSH
- proliferace buněk účinky léků fyziologie MeSH
- selenomethionin izolace a purifikace metabolismus MeSH
- techniky vsádkové kultivace metody MeSH
- velikost buňky účinky léků MeSH
- viabilita buněk účinky léků fyziologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- Publikační typ
- časopisecké články MeSH
The unicellular, nitrogen fixing cyanobacterium Cyanothece sp. ATCC 51142 is of a remarkable potential for production of third-generation biofuels. As the biotechnological potential of Cyanothece 51142 varies with the time of the day, we argue that it will, similarly, depend on the phase of the culture growth. Here, we study the batch culture dynamics to discover the dominant constraints in the individual growth phases and identify potential for inducing or delaying transitions between culture growth phases in Cyanothece 51142. We found that specific growth rate in the exponential phase of the culture is much less dependent on incident irradiance than the photosynthetic activity. We propose that surplus electrons that are released by water splitting are used in futile processes providing photoprotection additional to non-photochemical quenching. We confirm that the transition from exponential to linear phase is caused by a light limitation and the transition from linear to stationary phase by nitrogen limitation. We observe spontaneous diurnal metabolic oscillations in stationary phase culture that are synchronized over the entire culture without an external clue. We tentatively propose that the self-synchronization of the metabolic oscillations is due to a cell-to-cell communication of the cyanobacteria that is necessary for nitrogenase activity in nitrate depleted medium.
The effects of different volatile fatty acids (VFA, formate, acetate, propionate and butyrate), ammonium (NH (4) (+)) and agitation on methane (CH(4)) production were determined in 120-mL serum bottles. We showed that the addition of formate did not lead to an inhibition of methanogenesis until a concentration of 120 mmol/L. A complete inhibition of methanogenesis was detected in variants containing 360 mmol/L formate or propionate until day 3 but the production started afterwards within next 2 days. This might indicate a kind of adaptation to the higher volatile fatty acid concentrations. Increasing NH (4) (+) concentrations led to higher initial CH(4) production, with an optimum at 120 mmol/L. The addition of 720 mmol/L NH (4) (+) led to a complete inhibition until day 3; subsequently, CH(4) production started again on day 5 though it was still significantly lower compared to the other variants. Finally, also the speed of agitation showed significant effects on methanogenesis. The CH(4) production from complex carbon sources was most favourable at a moderate agitation of 150 rpm of the lab-scale serum bottles. A lower or higher speed brought about a distinct reduction of CH(4) production.
- MeSH
- Bacteria metabolismus MeSH
- biopaliva analýza mikrobiologie MeSH
- bioreaktory mikrobiologie MeSH
- kvartérní amoniové sloučeniny metabolismus MeSH
- kyseliny mastné těkavé metabolismus MeSH
- methan metabolismus MeSH
- odpadky - odstraňování MeSH
- odpadní vody analýza mikrobiologie MeSH
- rostliny metabolismus mikrobiologie MeSH
- techniky vsádkové kultivace přístrojové vybavení metody MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
