Reducing lactate accumulation has always been a goal of the mammalian cell biotechnology industry. When animal cells are cultured in vitro, the accumulation of lactate is mainly the combined result of two metabolic pathways. On one hand, glucose generates lactate under the function of lactate dehydrogenase A (LDHA); on the other hand, lactate can be oxidized to pyruvate by LDHB or LDHC and re-enter the TCA cycle. This study comprehensively evaluated the effects of LDH manipulation on the growth, metabolism and human adenovirus (HAdV) production of human embryonic kidney 293 (HEK-293) cells, providing a theoretical basis for engineering the lactate metabolism in mammalian cells. By knocking out ldha gene and overexpression of ldhb and ldhc genes, the metabolic efficiency of HEK-293 cells was effectively improved, and HAdV production was significantly increased. Compared with the control cell, LDH manipulation promoted cell growth, reduced the accumulation of lactate and ammonia, significantly enhanced the efficiency of substrate and energy metabolism of cells, and significantly increased the HAdV production capacity of HEK-293 cells. Among these LDH manipulation measures, ldhc gene overexpression performed the best, with the maximum cell density increased by about 38.7%. The yield of lactate to glucose and ammonia to glutamine decreased by 33.8% and 63.3%, respectively; and HAdV titer increased by at least 16 times. In addition, the ATP production rate, ATP/O₂ ratio, ATP/ADP ratio and NADH content of the modified cell lines were increased to varying degrees, and the energy metabolic efficiency was significantly improved.
Animals ; Humans ; L-Lactate Dehydrogenase/genetics* ; Lactic Acid ; Adenoviruses, Human ; Ammonia ; HEK293 Cells ; Glucose/metabolism* ; Adenosine Triphosphate/metabolism* ; Kidney/metabolism* ; Mammals/metabolism*

In the "Tutorial for outline of the healthy China 2030 plan", biomedicine was listed as a key planning and development area. Shanghai government also lists biomedicine as an emerging pillar industry. The rapid development of biomedicine industry put higher requirement for talents. Taking the idea of cross integration, mutually beneficial development, inheritance and innovation, the School of Biotechnology of East China University of Science and Technology organically integrates bioengineering and pharmaceutical majors to develop a new undergraduate engineering program of biomedicine, which specially reforms the talent training practice from the aspects of developing a "trinity teaching" standard system, a "three integration, three convergence" curriculum system, and a "three comprehensive education" innovative talent training system. We put forward the trinity of "value guidance, knowledge system, technology and non-technical core competence literacy" to foster emerging biomedicine engineering talents, and developed a comprehensive innovative talents training mode featured by "covering class-in and class-out, covering every student, and covering ideology and curriculum". Moreover, we established effective connections between courses and training goals, between general education courses and professional courses, and between top-notch talent training systems and training programs. Based on the achievements of teaching reform of the emerging engineering program "intelligent bio-manufacturing", the experience we obtained may provide ideas for development of the first-class bioengineering major in China.
Bioengineering ; Biomedical Engineering ; China ; Curriculum ; Humans ; Students

Microbiology is a key basic professional course for all the students specializing in biology, biotechnology and related majors. To date, microbiology is mainly taught in Chinese within colleges and universities in China. Development of a microbiology course that is taught in English may satisfy the diversified learning needs of the students and promote the "Double First-Class" initiative. We started to teach the microbiology course in English at the East China University of Science and Technology since 2016. This practice was associated with reform and innovation in the teaching methods and contents. The microbiology course taught in English greatly attracted the interest of the attending students and helped improve their professional English learning as well as scientific research. This course provided important support for fostering innovative professional first-class undergraduates under the context of the "Double First-Class" initiative.
China ; Humans ; Learning ; Students ; Universities

Currently, biomanufacturing technology and industry are receiving worldwide attention. However, there are still great challenges on bioprocess optimization and scale-up, including: lacing the process detection methods, which makes it difficult to meet the requirement of monitoring of key indicators and parameters; poor understanding of cell metabolism, which arouses problems to rationally achieve process optimization and regulation; the reactor environment is very different across the scales, resulting in low efficiency of stepwise scale-up. Considering the above key issues that need to be resolved, here we summarize the key technological innovations of the whole chain of fermentation process, i.e., real-time detection-dynamic regulation-rational scale-up, through case analysis. In the future, bioprocess design will be guided by a full lifecycle in-silico model integrating cellular physiology (spatiotemporal multiscale metabolic models) and fluid dynamics (CFD models). This will promote computer-aided design and development, accelerate the realization of large-scale intelligent production and serve to open a new era of green biomanufacturing.
Bioreactors ; Computer Simulation ; Fermentation ; Hydrodynamics

Starting from participating the high-level professional competition, our school has built a talent training system with the spirit of "biomaker" and an innovative practical ability training system. Such system takes the interest of student as the starting point, and relies on the strong scientific research and teaching infrastructure. The programme gives full play to students' initiatives and enhances the scientific research literacy and comprehensive ability of undergraduates majoring in biotechnology. It is an effective exploration of the traditional university education model and meets the urgent demand for innovative talents training in the era of rapid development of life sciences.
Biological Science Disciplines ; Biotechnology ; Genetic Engineering ; Humans ; Students ; Universities

Genome-scale metabolic network model (GSMM) is becoming an important tool for studying cellular metabolic characteristics, and remarkable advances in relevant theories and methods have been made. Recently, various constraint-based GSMMs that integrated genomic, transcriptomic, proteomic, and thermodynamic data have been developed. These developments, together with the theoretical breakthroughs, have greatly contributed to identification of target genes, systems metabolic engineering, drug discovery, understanding disease mechanism, and many others. This review summarizes how to incorporate transcriptomic, proteomic, and thermodynamic-constraints into GSMM, and illustrates the shortcomings and challenges of applying each of these methods. Finally, we illustrate how to develop and refine a fully integrated GSMM by incorporating transcriptomic, proteomic, and thermodynamic constraints, and discuss future perspectives of constraint-based GSMM.
Genome/genetics* ; Metabolic Engineering ; Metabolic Networks and Pathways/genetics* ; Models, Biological ; Proteomics

Bioreactors have been central in monoclonal antibodies and vaccines manufacturing by mammalian cells in suspension culture. Numerical simulation of five impeller combinations in a stirred bioreactor was conducted, and characteristics of velocity vectors, distributions of gas hold-up, distributions of shear rate in the bioreactor using 5 impeller combinations were numerically elucidated. In addition, genetically engineered CHO cells were cultivated in bioreactor installed with 5 different impeller combinations in fed-batch culture mode. The cell growth and antibody level were directly related to the maximum shear rate in the bioreactor, and the highest viable cell density and the peak antibody level were achieved in FBMI3 impeller combination, indicating that CHO cells are sensitive to shear force produced by impeller movement when cells were cultivated in bioreactor at large scale, and the maximum shear rate would play key roles in scaling-up of bioreactor at industrial scale.
Animals ; Batch Cell Culture Techniques ; Bioreactors ; standards ; CHO Cells ; Cell Count ; Computer Simulation ; Cricetinae ; Cricetulus ; Industrial Microbiology ; instrumentation ; methods

Objective:To explore the performance and clinical application value of a rapid detection method for the hematopoietic stem/progenitor cell of peripheral blood using the automated hematology analyzer.Methods:Methodology validation and retrospective study. Collected sample from Fujian Medical University Union Hospital from January 2015 to December 2018, the peripheral blood of 4 patients with acute myeloid leukemia was first treated, and one healthy donor′s peripheral blood stem cell collection 5 times diluent, for the methodology validation. And the peripheral venous blood and 5-fold dilutions of peripheral blood stem cell collection, from 23 patients with hematopoietic stem cell transplantation and 22 healthy donors of allogeneic peripheral blood stem cell transplantation, used for consistency retrospective analysis. In the linear test, each of the peripheral blood and HPC collecting solutions from blood cell separator, which known CD34 +cell concentration, that was high-value samples for the expected upper limit (H) . Another low-value sample is normal saline (L) . According to the multiple proportion dilution, HPC was detected and regressed consistency test specimens were 126, EDTA-K 2 anticoagulant venous blood 78 and peripheral blood stem cell 48. Venous blood was collected at the same time, one tube of blood routine and HPC detection, the other tube flow cytometry (FCM) detection of CD34 +cells. Stem cell collection was diluted 5 times with sterilized saline and divided into two tubes. One tube was used to count whole blood cells and HPC, the other tube was used to detect CD34 +cells by FCM. The test results of the two instruments were compared, and the deviation was evaluated. Results:The background counting was 0×10 9/L and the carryover rate was 0.1%, conformed to the quality requirements of hematology analyzer, and the repeatability study imprecision ranged between 4.7%-18.8%. HPC of peripheral venous blood linear range (0-27.201×10 9/L), Stem cell collection was diluted 5 times linear range (0-0.878×10 9/L). The results of 126 samples detected by the hematology analyzer and FCM were compared. The correlation coefficient r2=0.960 1. When WBC>10×10 9/L, the results of the two instruments have a good consistency. The slope is between 0.95 and 1.05, and the relative bias is less than 30%. Conclusions:This study suggests that the hematology analyzer has a good linear range for detecting HPC, and has a good correlation with FCM. The hematology analyzer has the advantages of no pretreatment, convenient operation, a wide range of applications in detecting HPC specimens.

Industrial bioprocess is a complex systematic process and bio-manufacturing can be realized on the basis of understanding the metabolism process of living cells. In this article, the multi-scale optimization principle and practice of industrial fermentation process are reviewed, including multi-scale optimizing theory and equipment, on-line sensing technology for cellular macroscopic metabolism, and correlated analysis of physiological parameters. Furthermore, intelligent control of industrial bioprocess is further addressed, in terms of new sensing technology for intracellular physiological metabolism, big database establishment and data depth calculation, intelligent decision.
Bioreactors ; Biotechnology ; Fermentation ; Industrial Microbiology

A rapid quantitative evaluation method for Siraitia grosvenorii cells was successfully developed based on plant cells' capacitance value detected by a viable cell mass monitor and the cryopreservation of S. grosvenorii suspension cells was optimized. The survival rate of S. grosvenorii cells was quantitatively measured by viable cell mass monitor and 2, 3, 5-triphenyltetrazolium chloride (TTC). An optimum cryoprotectant recipe is that the growth medium contained 10% sucrose and 10% DMSO. The experimental results also showed higher cell survival rates and cell viabilities were achieved when suspension cells were treated with pretreatment of 0.2 mol/L sucrose. With the increase of concentration of sucrose, however, the cell survival rate was decreased. And the cell survival rate represented a bell shape with the increase of pretreatment time. The highest cell survival rate and cell viability were obtained with the 9 h' s pretreatment. In addition, there was a good correlation between the cell survival rate measured by cell recovery test and that measured by viable cell mass monitor, while there were no significant differences in the cell morphology and the ability of mogrosides V production by S. grosvenorii cells cultured in suspension after cryopreservation. Therefore, the evaluation method developed based on the viable cell mass monitor has good feasibility and reliability.