Description and operation of a large-scale, mammalian cell, suspension culture facility

Abstract Gymnema sylvestre (Retz.) R.Br. ex Sm. is widely used as an efficient Ayurvedic traditional medicine for the treatment of diabetes. Phytochemical investigations of this plant showed gymnemic acids (a group of triterpenoid saponins) as the main active components. The present study aims to investigate the effectiveness of sodium nitroprusside (SNP) treatment for enhancement of cell suspension culture biomass and to evaluate their deacylgymnemic acid, gymnemagenin, gymnemic acid IV and gymnemic acid XVII contents of G. sylvestre. Callus was obtained from in vitro derived leaves of G. sylvestre on MS medium fortified with 3.0 mg/L 2, 4-D (2, 4-dichlorophenoxyacetic acid) and 2.0 mg/L Kn (Kinetin), and the same were used further to produce cell suspension cultures. Cell suspensions were exposed to different concentrations of SNP (5, 10, 20 and 40 µM) and data were collected at 20, 30 and 40 days. Out of the tested concentrations, 20 µM SNP had the highest level of cell culture growth (398.94 ± 8.32 g/L FCW and 40.00 ± 0.75 g/L DCW) on 40-day as compared to control. High-performance liquid chromatography analysis showed that maximum accumulation of deacylgymnemic acid (5.51 mg/g DCW), gymnemagenin (2.80 mg/g DCW) and gymnemic acid XVII (2.08 mg/g DCW) in 20 µM SNP treatment which is (13.43, 13.86 and17.33 folds) higher than the respective control at 40 days exposure. This research suggests that G. sylvestre cell suspension culture with optimal SNP elicitation treatment could be used as a good strategy for the large-scale production of these secondary metabolites at the industrial level.

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TINJAUAN Penggunaan Suspensi Sel dalam Kultur In Vitro

Jurnal AgroBiogen ◽

10.21082/jbio.v5n2.2009.p84-92 ◽

2016 ◽

Vol 5 (2) ◽

pp. 84 ◽

Cited By ~ 1

Author(s):

Sri Hutami

Keyword(s):

Secondary Metabolites ◽

Suspension Culture ◽

Cell Suspension ◽

Cell Suspension Culture ◽

Mass Production ◽

Large Scale ◽

Cell Suspension Cultures ◽

Suspension Cultures ◽

Cell Physiology

Cell suspension culture could be defined as a process that allows rapidly dividing homogenous suspension of cells to grow in liquid nutrient media. There are two main types of suspension cultures: (1) Batch cultures in which cells are nurtured in a fixed volume of medium until growth ceases and (2) Continuous cultures in which cell growth is maintained by continuous replenishment of sterile nutrient media. Plant cell suspension cultures are mostly used for the biochemical investigation of cell physiology, growth, metabolism, protoplast fusion, transformation and for large scale production of seed by bioreactor and production of secondary metabolites. Contamination is one of the largest problems when dealing with cell cultures. Differences between the products of cell suspension culture and whole plant are frequently observed. These phenomena’s may be resulted from lack of differentiation and organization and cell cultureinduced variation. Utilization of cell suspension culture in Indonesia is still limited, some of them for mass production of plantation seed with bioreactor system and for production of secondary metabolites. The success of this study give the opportunity for mass production of seeds from other plants and also production of secondary metabolites.

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Numerical simulation of scaling-up an inverted frusto-conical shaking bioreactor with low shear stress for mammalian cell suspension culture

Cytotechnology ◽

10.1007/s10616-019-00308-3 ◽

2019 ◽

Vol 71 (2) ◽

pp. 671-678

Author(s):

Ning Ding ◽

Chao Li ◽

Meijin Guo ◽

Ali Mohsin ◽

Siliang Zhang

Keyword(s):

Numerical Simulation ◽

Shear Stress ◽

Suspension Culture ◽

Cell Suspension ◽

Mammalian Cell ◽

Cell Suspension Culture ◽

Scaling Up ◽

Low Shear Stress ◽

Low Shear

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Computational fluid dynamics modeling of an inverted frustoconical shaking bioreactor for mammalian cell suspension culture

Biotechnology and Bioprocess Engineering ◽

10.1007/s12257-010-0426-z ◽

2011 ◽

Vol 16 (3) ◽

pp. 567-575 ◽

Cited By ~ 7

Author(s):

Haifeng Hang ◽

Yuanxin Guo ◽

Jian Liu ◽

Li Bai ◽

Jianye Xia ◽

...

Keyword(s):

Fluid Dynamics ◽

Computational Fluid Dynamics ◽

Suspension Culture ◽

Cell Suspension ◽

Mammalian Cell ◽

Cell Suspension Culture ◽

Dynamics Modeling ◽

Computational Fluid Dynamics Modeling

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Engineering Considerations to Produce Bioactive Compounds from Plant Cell Suspension Culture in Bioreactors

Plants ◽

10.3390/plants10122762 ◽

2021 ◽

Vol 10 (12) ◽

pp. 2762

Author(s):

Elizabeth Alejandra Motolinía-Alcántara ◽

Carlos Omar Castillo-Araiza ◽

Mario Rodríguez-Monroy ◽

Angélica Román-Guerrero ◽

Francisco Cruz-Sosa

Keyword(s):

Suspension Culture ◽

Cell Suspension ◽

Cell Suspension Culture ◽

Large Scale ◽

Scaling Up ◽

Operating Conditions ◽

Scale Production ◽

Plant Cell Suspension Culture ◽

Plant Cell Suspension ◽

Large Scale Production

The large-scale production of plant-derived secondary metabolites (PDSM) in bioreactors to meet the increasing demand for bioactive compounds for the treatment and prevention of degenerative diseases is nowadays considered an engineering challenge due to the large number of operational factors that need to be considered during their design and scale-up. The plant cell suspension culture (CSC) has presented numerous benefits over other technologies, such as the conventional whole-plant extraction, not only for avoiding the overexploitation of plant species, but also for achieving better yields and having excellent scaling-up attributes. The selection of the bioreactor configuration depends on intrinsic cell culture properties and engineering considerations related to the effect of operating conditions on thermodynamics, kinetics, and transport phenomena, which together are essential for accomplishing the large-scale production of PDSM. To this end, this review, firstly, provides a comprehensive appraisement of PDSM, essentially those with demonstrated importance and utilization in pharmaceutical industries. Then, special attention is given to PDSM obtained out of CSC. Finally, engineering aspects related to the bioreactor configuration for CSC stating the effect of the operating conditions on kinetics and transport phenomena and, hence, on the cell viability and production of PDSM are presented accordingly. The engineering analysis of the reviewed bioreactor configurations for CSC will pave the way for future research focused on their scaling up, to produce high value-added PDSM.

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