Application of modified Logistic and Monod models in a single equations system framework to improve cell culture growth modeling and estimation

In modeling cell culture growth using unstructured model, two types of equations are normally used: logistic and Monod. However, these two equations are known for their limitations to model death phase of cell culture growth and to account for dead cells accumulation data. In this paper, we present a modeling framework whereby both Logistic and Monod equations can be used in a single set of equations system to overcome these limitations. First, it can be shown that the increase of total cell population that consists of viable and dead cells follows a logistic growth pattern with its own intrinsic growth rate and total carrying capacity. Furthermore, a hybrid Logistic-Monod equation with first-order decay kinetics can be used to model viable cell growth data with decline phase effectively. With this paradigm, a pseudo-rate equation can be written to account for dead cells accumulation data using population balancing with a simple understanding that dead cell population is simply the difference between total and viable cells. These equations can be adjoined with substrate consumption and product generation rate equations to depict complete batch growth data that covers exponential growth and death phases. This modeling framework has been fitted successfully to fit batch growth data of two cell lines from published literature with complete depictions of dead cell accumulation and cell viability profiles. The implication of this modeling framework for chemostat culture performance analysis is further investigated.

Applying Logistic and Monod models in a single equations system framework for cell culture growth modeling and estimation

In modeling cell culture growth, two types of modeling equations are normally used: logistic and Monod. These two equations are known for their strengths and weaknesses in modeling cell culture growth. In this contribution, we show how these equations can be used in a single equations system framework to model cell culture growth that is supported by experimental observation. Specifically, we propose that logistic equation is used to model the dynamic of total cells growth that is simply the summation of viable and dead cells populations in the system. Subsequently, Monod equation is used to model the dynamic of viable cells growth that is subjected to growth-limiting substrate and cells death rate term. With this paradigm, a rate equation can be written for the accumulation of dead cells in the system with a simple understanding that dead cells population is simply the difference between total and viable cells. These equations can be adjoined with appropriate substrate consumption and product generation rate equations to depict a complete time course profiles of batch culture experiment. This modeling framework has been fitted successfully to depict a batch growth data of IgG-secreting murine hybridoma cell from published literature.

Applying Logistic and Monod models in a single equations system framework for cell culture growth modeling and estimation

In modeling cell culture growth, two types of modeling equations are normally used: logistic and Monod. These two equations are known for their strengths and weaknesses in modeling cell culture growth. In this contribution, we show how these equations can be used in a single equations system framework to model cell culture growth that is supported by experimental observation. Specifically, we propose that logistic equation is used to model the dynamic of total cells growth that is simply the summation of viable and dead cells populations in the system. Subsequently, Monod equation is used to model the dynamic of viable cells growth that is subjected to growth-limiting substrate and cells death rate term. With this paradigm, a rate equation can be written for the accumulation of dead cells in the system with a simple understanding that dead cells population is simply the difference between total and viable cells. These equations can be adjoined with appropriate substrate consumption and product generation rate equations to depict a complete time course profiles of batch culture experiment. This modeling framework has been fitted successfully to depict a batch growth data of IgG-secreting murine hybridoma cell from published literature.

In-vitro assessment of appropriate hydrophilic scaffolds by co-electrospinning of poly(1,4 cyclohexane isosorbide terephthalate)/polyvinyl alcohol

Textile-based Scaffolds preparation has the attractive features to fulfill the stated and implied needs of the consumer but there are still challenges of stability, elongation, appreciable bio-compatibility, and stated hydrophilic behavior. To overcome these challenges, the authors tried to fabricate a scaffold by blending of two highly biocompatible polymers; polyvinyl alcohol and poly(1,4 cyclohexane isosorbide terephthalate) through co-electrospinning. The resultant scaffold by the stated innovative approach evaluated from different characterizations such as dimensional stability/morphology was evaluated by scanning electron microscopy, chemical interactions by that Fourier transmission infrared spectra, wetting behavior was analyzed by a static angle with a contact angle meter from drop method, elongation was examined by tensile strength tester and in-vitro assessment was done by MTT analysis. Based on verified results, it was concluded that PVA/PICT scaffold has a potential for dual nature of hydrophilicity & hydrophobicity and appreciable cell culture growth, stated dimensional stability and suitable elongation as per requirements of the nature of scaffold.

The Effect of Single Co-expression of The DnaK-DnaJ-GrpE and GroEL/ES Chaperones and Their Combination on Expression Intein-pretrombin-2 in Escherichia coli ER2566

The use of recombinant thrombin in the manufacture of fibrin glue allows diseases contamination to be avoided. However, the expression of recombinant protein in E. coli still has a disadvantage of the formation of inclusion bodies, so it needs to be minimized by co-expression of chaperones. Therefore, the aim of this study was to determine the effect of single DnaK-DnaJ-GrpE and GroEL/ES chaperone expression and their combination on the expression of intein-pretrombin-2Ti,pH on E. coli ER2566. The method started with isolation of pTWIN1-prethrombin-2Ti,pH and pG-KJE8 from E. coli TOP10F' and DH5α respectively, the co-transformation of the expression host E. coli ER2566 using pG-KJE8 and pTWIN1-prethrombin-2Ti,pHvectors, the chaperone co-expression was induced using L-Arabinosa before IPTG induction and cell culture growth was incubated at 22 oC. The expression products were characterized by using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. The results of the co-expression of chaperone showed that the number of soluble fraction was higher than the one without co-expression of chaperone. In addition, the co-expression of chaperone using pG-KJE8 in intein-prothrombin-2Ti,pH expression was sufficient using tetracycline as an inducer.

Cyclodextrin-Elicited Bryophyllum Suspension Cultured Cells: Enhancement of the Production of Bioactive Compounds

The rates of production of secondary metabolites obtained by employing conventional plant breeding may be low for practical purposes. Thus, innovative approaches for increasing their rates of production are being developed. Here, we propose the use of elicited plant suspension cultured cells (PSCC) with cyclodextrins (CDs) as an alternative method for the production of bioactive compounds from Bryophyllum species. For this purpose, we analyzed the effects of methyl–β–cyclodextrin and 2–hydroxypropyl–β–cyclodextrin on cell culture growth and on the intra- and extracellular production of phenols and flavonoids. Results clearly show that CDs enhance the biosynthesis of polyphenols by PSCC favoring their accumulation outside the cells. CDs shift the homeostatic equilibrium by complexing extracellular phenolics, causing stress in cells that respond by increasing the production of intracellular phenolics. We also analyzed the radical scavenging activity of the culture medium extracts against 2,2–diphenyl–1–pycrilhydrazyl (DPPH) radical, which increased with respect to the control samples (no added CDs). Our results suggest that both the increase in the production of polyphenols and their radical scavenging activity are a consequence of their inclusion in the CD cavities. Overall, based on our findings, CDs can be employed as hosts for increasing the production of polyphenols from Bryophyllum species.

Cyclodextrin-elicited Bryophyllum Suspension Cultured Cells: Enhancement of the Production of Bioactive Compounds

The rates of production of secondary metabolites obtained by employing conventional plant breeding may be low for practical purposes. Thus, innovative approaches for increasing their rates of production are being developed. Here, we propose the use of elicited suspension plant cultured cells (PSCC) with cyclodextrins (CDs) as an alternative method for the production of bioactive compounds from Bryophyllum species. For the purpose, we analyzed the effects of methyl--cyclodextrin and 2-hydroxypropyl--cyclodextrin on cell culture growth and on the intra- and extracellular production of phenols and flavonoids. Results clearly show that CDs enhance the biosynthesis of polyphenols by PSCC favoring their accumulation outside the cells. CDs shift the homeostatic equilibrium by complexing extracellular phenolics, causing stress in cells that respond by increasing the production of intracellular phenolics. We also analyzed the radical scavenging activity of the culture medium extracts against DPPH• radical, which increased with respect to the control samples (no added CDs). Our results suggest that both the increase in the production of polyphenols and their radical scavenging activity are a consequence of their inclusion in the CD cavities. Overall, based on our findings, CDs can be employed as hosts for increasing the production of polyphenols from Bryophyllum species.