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Stem Cell Research & Therapy Jun 2023Induced pluripotent stem cells (iPSCs) offer potential to revolutionize regenerative medicine as a renewable source for islets, dopaminergic neurons, retinal cells, and...
BACKGROUND
Induced pluripotent stem cells (iPSCs) offer potential to revolutionize regenerative medicine as a renewable source for islets, dopaminergic neurons, retinal cells, and cardiomyocytes. However, translation of these regenerative cell therapies requires cost-efficient mass manufacturing of high-quality human iPSCs. This study presents an improved three-dimensional Vertical-Wheel® bioreactor (3D suspension) cell expansion protocol with comparison to a two-dimensional (2D planar) protocol.
METHODS
Sendai virus transfection of human peripheral blood mononuclear cells was used to establish mycoplasma and virus free iPSC lines without common genetic duplications or deletions. iPSCs were then expanded under 2D planar and 3D suspension culture conditions. We comparatively evaluated cell expansion capacity, genetic integrity, pluripotency phenotype, and in vitro and in vivo pluripotency potential of iPSCs.
RESULTS
Expansion of iPSCs using Vertical-Wheel® bioreactors achieved 93.8-fold (IQR 30.2) growth compared to 19.1 (IQR 4.0) in 2D (p < 0.0022), the largest expansion potential reported to date over 5 days. 0.5 L Vertical-Wheel® bioreactors achieved similar expansion and further reduced iPSC production cost. 3D suspension expanded cells had increased proliferation, measured as Ki67 expression using flow cytometry (3D: 69.4% [IQR 5.5%] vs. 2D: 57.4% [IQR 10.9%], p = 0.0022), and had a higher frequency of pluripotency marker (Oct4NanogSox2) expression (3D: 94.3 [IQR 1.4] vs. 2D: 52.5% [IQR 5.6], p = 0.0079). q-PCR genetic analysis demonstrated a lack of duplications or deletions at the 8 most commonly mutated regions within iPSC lines after long-term passaging (> 25). 2D-cultured cells displayed a primed pluripotency phenotype, which transitioned to naïve after 3D-culture. Both 2D and 3D cells were capable of trilineage differentiation and following teratoma, 2D-expanded cells generated predominantly solid teratomas, while 3D-expanded cells produced more mature and predominantly cystic teratomas with lower Ki67 expression within teratomas (3D: 16.7% [IQR 3.2%] vs.. 2D: 45.3% [IQR 3.0%], p = 0.002) in keeping with a naïve phenotype.
CONCLUSION
This study demonstrates nearly 100-fold iPSC expansion over 5-days using our 3D suspension culture protocol in Vertical-Wheel® bioreactors, the largest cell growth reported to date. 3D expanded cells showed enhanced in vitro and in vivo pluripotency phenotype that may support more efficient scale-up strategies and safer clinical implementation.
Topics: Humans; Induced Pluripotent Stem Cells; Ki-67 Antigen; Leukocytes, Mononuclear; Cell Differentiation; Phenotype; Teratoma
PubMed: 37280707
DOI: 10.1186/s13287-023-03382-9 -
International Journal of Molecular... Jul 2023An organoid is a 3D organization of cells that can recapitulate some of the structure and function of native tissue. Recent work has seen organoids gain prominence as a... (Review)
Review
An organoid is a 3D organization of cells that can recapitulate some of the structure and function of native tissue. Recent work has seen organoids gain prominence as a valuable model for studying tissue development, drug discovery, and potential clinical applications. The requirements for the successful culture of organoids in vitro differ significantly from those of traditional monolayer cell cultures. The generation and maturation of high-fidelity organoids entails developing and optimizing environmental conditions to provide the optimal cues for growth and 3D maturation, such as oxygenation, mechanical and fluidic activation, nutrition gradients, etc. To this end, we discuss the four main categories of bioreactors used for organoid culture: stirred bioreactors (SBR), microfluidic bioreactors (MFB), rotating wall vessels (RWV), and electrically stimulating (ES) bioreactors. We aim to lay out the state-of-the-art of both commercial and in-house developed bioreactor systems, their benefits to the culture of organoids derived from various cells and tissues, and the limitations of bioreactor technology, including sterilization, accessibility, and suitability and ease of use for long-term culture. Finally, we discuss future directions for improvements to existing bioreactor technology and how they may be used to enhance organoid culture for specific applications.
Topics: Cell Culture Techniques; Organoids; Bioreactors
PubMed: 37511186
DOI: 10.3390/ijms241411427 -
Frontiers in Plant Science 2023Ever since agriculture started, plants have been bred to obtain better yields, better fruits, or sustainable products under uncertain biotic and abiotic conditions.... (Review)
Review
Ever since agriculture started, plants have been bred to obtain better yields, better fruits, or sustainable products under uncertain biotic and abiotic conditions. However, a new way to obtain products from plant cells emerged with the development of recombinant DNA technologies. This led to the possibility of producing exogenous molecules in plants. Furthermore, plant chemodiversity has been the main source of pharmacological molecules, opening a field of plant biotechnology directed to produce high quality plant metabolites. The need for different products by the pharma, cosmetics agriculture and food industry has pushed again to develop new procedures. These include cell production in bioreactors. While plant tissue and cell culture are an established technology, beginning over a hundred years ago, plant cell cultures have shown little impact in biotechnology projects, compared to bacterial, yeasts or animal cells. In this review we address the different types of bioreactors that are currently used for plant cell production and their usage for quality biomolecule production. We make an overview of , , , , and as well-established models for plant cell culture, and some species used to obtain important metabolites, with an insight into the type of bioreactor and production protocols.
PubMed: 38148861
DOI: 10.3389/fpls.2023.1310405 -
Stem Cell Research & Therapy Oct 2023Human umbilical cord mesenchymal stem cells (hUC-MSCs) are widely used in cell therapy due to their robust immunomodulatory and tissue regenerative capabilities....
BACKGROUND
Human umbilical cord mesenchymal stem cells (hUC-MSCs) are widely used in cell therapy due to their robust immunomodulatory and tissue regenerative capabilities. Currently, the predominant method for obtaining hUC-MSCs for clinical use is through planar culture expansion, which presents several limitations. Specifically, continuous cell passaging can lead to cellular aging, susceptibility to contamination, and an absence of process monitoring and control, among other limitations. To overcome these challenges, the technology of microcarrier-bioreactor culture was developed with the aim of ensuring the therapeutic efficacy of cells while enabling large-scale expansion to meet clinical requirements. However, there is still a knowledge gap regarding the comparison of biological differences in cells obtained through different culture methods.
METHODS
We developed a culture process for hUC-MSCs using self-made microcarrier and stirred bioreactor. This study systematically compares the biological properties of hUC-MSCs amplified through planar culture and microcarrier-bioreactor systems. Additionally, RNA-seq was employed to compare the differences in gene expression profiles between the two cultures, facilitating the identification of pathways and genes associated with cell aging.
RESULTS
The findings revealed that hUC-MSCs expanded on microcarriers exhibited a lower degree of cellular aging compared to those expanded through planar culture. Additionally, these microcarrier-expanded hUC-MSCs showed an enhanced proliferation capacity and a reduced number of cells in the cell cycle retardation period. Moreover, bioreactor-cultured cells differ significantly from planar cultures in the expression of genes associated with the cytoskeleton and extracellular matrix.
CONCLUSIONS
The results of this study demonstrate that our microcarrier-bioreactor culture method enhances the proliferation efficiency of hUC-MSCs. Moreover, this culture method exhibits the potential to delay the process of cell aging while preserving the essential stem cell properties of hUC-MSCs.
Topics: Humans; Mesenchymal Stem Cells; Cells, Cultured; Cellular Senescence; Stem Cells; Bioreactors; Umbilical Cord; Cell Differentiation; Mesenchymal Stem Cell Transplantation
PubMed: 37794520
DOI: 10.1186/s13287-023-03514-1 -
World Journal of Microbiology &... Dec 2023Increased production of renewable energy sources is becoming increasingly needed. Amidst other strategies, one promising technology that could help achieve this goal is... (Review)
Review
Increased production of renewable energy sources is becoming increasingly needed. Amidst other strategies, one promising technology that could help achieve this goal is biological hydrogen production. This technology uses micro-organisms to convert organic matter into hydrogen gas, a clean and versatile fuel that can be used in a wide range of applications. While biohydrogen production is in its early stages, several challenges must be addressed for biological hydrogen production to become a viable commercial solution. From an experimental perspective, the need to improve the efficiency of hydrogen production, the optimization strategy of the microbial consortia, and the reduction in costs associated with the process is still required. From a scale-up perspective, novel strategies (such as modelling and experimental validation) need to be discussed to facilitate this hydrogen production process. Hence, this review considers hydrogen production, not within the framework of a particular production method or technique, but rather outlines the work (bioreactor modes and configurations, modelling, and techno-economic and life cycle assessment) that has been done in the field as a whole. This type of analysis allows for the abstraction of the biohydrogen production technology industrially, giving insights into novel applications, cross-pollination of separate lines of inquiry, and giving a reference point for researchers and industrial developers in the field of biohydrogen production.
Topics: Fermentation; Bioreactors; Microbial Consortia; Hydrogen; Costs and Cost Analysis; Biofuels
PubMed: 38057658
DOI: 10.1007/s11274-023-03845-4 -
Molecules (Basel, Switzerland) Apr 2024Bioremediation uses the degradation abilities of microorganisms and other organisms to remove harmful pollutants that pollute the natural environment, helping return it... (Review)
Review
Bioremediation uses the degradation abilities of microorganisms and other organisms to remove harmful pollutants that pollute the natural environment, helping return it to a natural state that is free of harmful substances. Organism-derived enzymes can degrade and eliminate a variety of pollutants and transform them into non-toxic forms; as such, they are expected to be used in bioremediation. However, since enzymes are proteins, the low operational stability and catalytic efficiency of free enzyme-based degradation systems need improvement. Enzyme immobilization methods are often used to overcome these challenges. Several enzyme immobilization methods have been applied to improve operational stability and reduce remediation costs. Herein, we review recent advancements in immobilized enzymes for bioremediation and summarize the methods for preparing immobilized enzymes for use as catalysts and in pollutant degradation systems. Additionally, the advantages, limitations, and future perspectives of immobilized enzymes in bioremediation are discussed.
Topics: Biodegradation, Environmental; Enzymes, Immobilized; Environmental Pollutants; Bioreactors; Hazardous Substances
PubMed: 38731512
DOI: 10.3390/molecules29092021 -
Applied Microbiology and Biotechnology Jul 2023Stem cell-based cell therapeutics and especially those based on human mesenchymal stem cells (hMSCs) and induced pluripotent stem cells (hiPSCs) are said to have... (Review)
Review
Stem cell-based cell therapeutics and especially those based on human mesenchymal stem cells (hMSCs) and induced pluripotent stem cells (hiPSCs) are said to have enormous developmental potential in the coming years. Their applications range from the treatment of orthopedic disorders and cardiovascular diseases to autoimmune diseases and even cancer. However, while more than 27 hMSC-derived therapeutics are currently commercially available, hiPSC-based therapeutics have yet to complete the regulatory approval process. Based on a review of the current commercially available hMSC-derived therapeutic products and upcoming hiPSC-derived products in phase 2 and 3, this paper compares the cell therapy manufacturing process between these two cell types. Moreover, the similarities as well as differences are highlighted and the resulting impact on the production process discussed. Here, emphasis is placed on (i) hMSC and hiPSC characteristics, safety, and ethical aspects, (ii) their morphology and process requirements, as well as (iii) their 2- and 3-dimensional cultivations in dependence of the applied culture medium and process mode. In doing so, also downstream processing aspects are covered and the role of single-use technology is discussed. KEY POINTS: • Mesenchymal and induced pluripotent stem cells exhibit distinct behaviors during cultivation • Single-use stirred bioreactor systems are preferred for the cultivation of both cell types • Future research should adapt and modify downstream processes to available single-use devices.
Topics: Humans; Induced Pluripotent Stem Cells; Cell Culture Techniques; Cell- and Tissue-Based Therapy; Culture Media; Bioreactors; Cell Differentiation
PubMed: 37246986
DOI: 10.1007/s00253-023-12583-4 -
Chemosphere Dec 2023Nowadays, people spend 80-90% of their time indoors, while recent policies on energy efficient and safe buildings require reduced building ventilation rates and locked... (Review)
Review
Nowadays, people spend 80-90% of their time indoors, while recent policies on energy efficient and safe buildings require reduced building ventilation rates and locked windows. These facts have raised a growing concern on indoor air quality, which is currently receiving even more attention than outdoors pollution. Prevention is the first and most cost-effective strategy to improve indoor air quality, but once pollution is generated, a battery of physicochemical technologies is typically implemented to improve air quality with a questionable efficiency and at high operating costs. Biotechnologies have emerged as promising alternatives to abate indoor air pollutants, but current bioreactor configurations and the low concentrations of indoor air pollutants limit their widespread implementation in homes, offices and public buildings. In this context, recent investigations have shown that potted plants can aid in the removal of a wide range of indoor air pollutants, especially volatile organic compounds (VOCs), and can be engineered in aesthetically attractive configurations. The original investigations conducted by NASA, along with recent advances in technology and design, have resulted in a new generation of botanical biofilters with the potential to effectively mitigate indoor air pollution, with increasing public aesthetics acceptance. This article presents a review of the research on active botanical filters as sustainable alternatives to purify indoor air.
Topics: Humans; Air Pollutants; Air Pollution, Indoor; Bioreactors; Biotechnology; Electric Power Supplies; Volatile Organic Compounds
PubMed: 37863205
DOI: 10.1016/j.chemosphere.2023.140483 -
Foods (Basel, Switzerland) Dec 2023Pathogenic biofilm formation within food processing industries raises a serious public health and safety concern, and places burdens on the economy. Biofilm formation on... (Review)
Review
Pathogenic biofilm formation within food processing industries raises a serious public health and safety concern, and places burdens on the economy. Biofilm formation on equipment surfaces is a rather complex phenomenon, wherein multiple steps are involved in bacterial biofilm formation. In this review we discuss the stages of biofilm formation, the existing literature on the impact of surface properties and shear stress on biofilms, types of bioreactors, and antimicrobial coatings. The review underscores the significance of prioritizing biofilm prevention strategies as a first line of defense, followed by control measures. Utilizing specific biofilm eradication strategies as opposed to a uniform approach is crucial because biofilms exhibit different behavioral outcomes even amongst the same species when the environmental conditions change. This review is geared towards biofilm researchers and food safety experts, and seeks to derive insights into the scope of biofilm formation, prevention, and control. The use of suitable bioreactors is paramount to understanding the mechanisms of biofilm formation. The findings provide useful information to researchers involved in bioreactor selection for biofilm investigation, and food processors in surfaces with novel antimicrobial coatings, which provide minimal bacterial attachment.
PubMed: 38137299
DOI: 10.3390/foods12244495 -
Journal of Environmental Management May 2024This study focused on the economic feasibility of two potential industrial-scale bioleaching technologies for metal recovery from specific metallurgical by-products,...
This study focused on the economic feasibility of two potential industrial-scale bioleaching technologies for metal recovery from specific metallurgical by-products, mainly basic oxygen steelmaking dust (BOS-D) and goethite. The investigation compared two bioleaching scaling technology configurations, including an aerated bioreactor and an aerated and stirred bioreactor across different scenarios. Results indicated that bioleaching using Acidithiobacillus ferrooxidans proved financially viable for copper extraction from goethite, particularly when 5% and 10% pulp densities were used in the aerated bioreactor, and when 10% pulp density was used in the aerated and stirred bioreactor. Notably, a net present value (NPV) of $1,275,499k and an internal rate of return (IRR) of 65% for Cu recovery from goethite were achieved over 20-years after project started using the aerated and stirred bioreactor plant with a capital expenditure (CAPEX) of $119,816,550 and an operational expenditure (OPEX) of $5,896,580/year. It is expected that plant will start to make profit after one year of operation. Aerated and stirred bioreactor plant appeared more reliable alternative compared to the aerated bioreactor plant as the plant consists of 12 reactors which can allow better management and operation in small volume with multiple reactors. Despite the limitations, this techno-economic assessment emphasized the significance of selective metal recovery and plant design, and underscored the major expenses associated with the process.
Topics: Bioreactors; Metallurgy; Acidithiobacillus; Copper; Minerals; Iron Compounds
PubMed: 38643624
DOI: 10.1016/j.jenvman.2024.120904