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Optimization of co-culture inoculated microbial fuel cell performance using response surface methodology

Islam MA, Ong HR, Ethiraj B, Cheng CK, Rahman Khan MM

J Environ Manage, 2018 Nov 01;225:242-251.
PMID: 30092551 DOI: 10.1016/j.jenvman.2018.08.002

Microbial fuel cells (MFCs) are considered as promising technology to achieve simultaneous wastewater treatment and electricity generation. However, operational and technological developments are still required to make it as a sustainable technology. In the present study, response surface methodology (RSM) was used to evaluate the effects of substrate concentration, co-culture composition, pH and time on the performance of co-culture (Klebsiella variicola and Pseudomonas aeruginosa) inoculated double chamber MFC. From the statistical analysis, it can be seen that the performance of MFC was not influenced by the interaction between the initial COD and time, pH and time, pH and initial COD, time and initial COD. However, the interaction between the inoculum composition and time, pH and the inoculum composition, initial COD and inoculum composition significantly influenced the performance of MFC. Based on the RSM results, best performance (power density and COD removal efficiency) was obtained when the inoculum composition, initial COD, pH and time were about 1:1, 26.690 mg/L, 7.21 and 15.50 days, respectively. The predictions from the model were in close agreement with the experimental results suggesting that the proposed model could adequately represent the actual relationships between the independent variables generating electricity and the COD removal efficiency.

Matched MeSH terms: Coculture Techniques*
Microbial synergistic interactions enhanced power generation in co-culture driven microbial fuel cell

Islam MA, Karim A, Mishra P, Dubowski JJ, Yousuf A, Sarmin S, et al.

Sci Total Environ, 2020 Oct 10;738:140138.
PMID: 32806344 DOI: 10.1016/j.scitotenv.2020.140138

An understanding of the inter-species relationships, especially their metabolic network in a mixed-culture system, is crucial to design an effective inoculum for enhancing the power generation of wastewater fed microbial fuel cell (MFC). In the present study, the influence of microbial mutualistic interactions on the power generation of palm oil mill effluent fed MFCs has been widely investigated by designing several co-culture and mixed culture inoculums. Among the different inoculum compositions, the highest power density of 14.8 W/m3 was achieved by Pseudomonas aeruginosa and Klebsiella variicola co-culture inoculum due to their synergistic relationships which were inter-linked via fermentation-based metabolites. Besides, the interaction of K. variicola and Bacillus cereus positively influenced the power generation resulting in a maximum power density of 11.8 W/m3 whereas the antagonistic relationship between B. cereus and P. aeruginosa resulted in a lower power generation of 1.9 W/m3. The microbial mutualistic interactions were investigated with polarization, cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), as well as by using metabolite and biofilm analysis. It was observed that the synergism between bacteria enhanced power generation through the production of higher electron shuttling mediators and efficient biofilm formation as evidenced by polarization, CV and EIS analysis. In contrast, the antagonistic relationship resulted in production of cell inhibiting metabolites leading to the formation of ineffective biofilm. These findings demonstrate that the synergistic interaction between or within microorganisms is emergent in designing co-culture or mixed-culture inoculum for achieving maximum power generation in MFCs.

Matched MeSH terms: Coculture Techniques
A review on microalgal-bacterial co-culture: The multifaceted role of beneficial bacteria towards enhancement of microalgal metabolite production

Tong CY, Honda K, Derek CJC

Environ Res, 2023 Jul 01;228:115872.
PMID: 37054838 DOI: 10.1016/j.envres.2023.115872

Mass microalgal-bacterial co-cultures have come to the fore of applied physiological research, in particularly for the optimization of high-value metabolite from microalgae. These co-cultures rely on the existence of a phycosphere which harbors unique cross-kingdom associations that are a prerequisite for the cooperative interactions. However, detailed mechanisms underpinning the beneficial bacterial effects onto microalgal growth and metabolic production are rather limited at the moment. Hence, the main purpose of this review is to shed light on how bacteria fuels microalgal metabolism or vice versa during mutualistic interactions, building upon the phycosphere which is a hotspot for chemical exchange. Nutrients exchange and signal transduction between two not only increase the algal productivity, but also facilitate in the degradation of bio-products and elevate the host defense ability. Main chemical mediators such as photosynthetic oxygen, N-acyl-homoserine lactone, siderophore and vitamin B12 were identified to elucidate beneficial cascading effects from the bacteria towards microalgal metabolites. In terms of applications, the enhancement of soluble microalgal metabolites is often associated with bacteria-mediated cell autolysis while bacterial bio-flocculants can aid in microalgal biomass harvesting. In addition, this review goes in depth into the discussion on enzyme-based communication via metabolic engineering such as gene modification, cellular metabolic pathway fine-tuning, over expression of target enzymes, and diversion of flux toward key metabolites. Furthermore, possible challenges and recommendations aimed at stimulating microalgal metabolite production are outlined. As more evidence emerges regarding the multifaceted role of beneficial bacteria, it will be crucial to incorporate these findings into the development of algal biotechnology.

Matched MeSH terms: Coculture Techniques
Enhancing food preservation with postbiotic metabolites γ-aminobutyric acid (GABA) and bacteriocin-like inhibitory substances (BLIS) produced by Lactobacillus brevis C23 co-cultures in plant-based medium

Chuah WW, Tan JS, Hazwani Oslan SN, Bothi Raja P

Prep Biochem Biotechnol, 2024 Apr;54(4):514-525.
PMID: 37694843 DOI: 10.1080/10826068.2023.2252047

Lactic acid bacteria (LAB) can produce γ-aminobutyric acid (GABA) with antioxidant properties and sedative effects when it binds to the GABA receptor in the human brain. LAB can also produce bacteriocin-like inhibitory substances (BLIS) with antimicrobial capabilities during carbohydrate fermentation. GABA and BLIS are natural compounds with potential health benefits and food preservation properties. Lactobacillus brevis C23 was co-cultured with three different LABs as inducers, which produced the highest GABA content and BLIS activity. They were cultured in various plant-based media to obtain an edible and better-tasting final product over commercially available media like MRS broth. A coconut-based medium with additives was optimized using response surface methodology (RSM) to increase GABA and BLIS production. The optimized medium for maximum GABA production (3.22 ± 0.01 mg/mL) and BLIS activity (84.40 ± 0.44%) was a 5.5% coconut medium containing 0.23% glucose, 1.44% Tween 20, 0.48% L-glutamic acid, and 0.02% pyridoxine. Due to the presence of GABA, the cell-free supernatant (CFS) as a postbiotic showed higher antioxidant activity than other food preservatives like nisin and potassium sorbate. Finally, microbiological tests on food samples showed that the postbiotic was more effective than other preservatives at combating the growth of LAB, molds and coliform bacteria, making it a possible food preservative.

Matched MeSH terms: Coculture Techniques
Fulltext Marine microalgae co-cultured with floc-forming bacterium: Insight into growth and lipid productivity

Yee CS, Okomoda VT, Hashim F, Waiho K, Sheikh Abdullah SR, Alamanjo C, et al.

PeerJ, 2021;9:e11217.
PMID: 33981498 DOI: 10.7717/peerj.11217

This study investigated the effect of co-culturing microalgae with a floc-forming bacterium. Of the six microalgae isolated from a biofloc sample, only Thalassiosira weissflogii, Chlamydomonas sp. and Chlorella vulgaris were propagated successfully in Conway medium. Hence, these species were selected for the experiment comparing microalgae axenic culture and co-culture with the floc-forming bacterium, Bacillus infantis. Results obtained showed that the co-culture had higher microalgae biomass compared to the axenic culture. A similar trend was also observed concerning the lipid content of the microalgae-bacterium co-cultures. The cell number of B. infantis co-cultured with T. weissflogii increased during the exponential stage until the sixth day, but the other microalgae species experienced a significant early reduction in cell density of the bacteria at the exponential stage. This study represents the first attempt at co-culturing microalgae with B. infantis, a floc-forming bacterium, and observed increased biomass growth and lipid accumulation compared to the axenic culture.

Matched MeSH terms: Coculture Techniques
Multi-objectives optimization on microwave-assisted-biological-based biogas upgrading and bio-succinic acid production

Adnan AI, Ong MY, Mohamed H, Chia SR, Milano J, Nomanbhay S

Bioresour Technol, 2024 Aug;406:131028.
PMID: 38914237 DOI: 10.1016/j.biortech.2024.131028

This study represents the first investigation of bio-succinic acid (bio-SA) production with methane enrichment using carbon-dioxide-fixating bacteria in the co-culture of ragi tapai and macroalgae, Chaetomorpha. Microwave irradiation has also been introduced to enhance the biochemical processes as it could provide rapid and selective heating of substrates. In this research, microwave irradiation was applied on ragi tapai as a pre-treatment process. Factors such as microwave irradiation dose on ragi tapai, Chaetomorpha ratio in the co-culture, and pH value were studied. Optimal conditions were identified using Design-Expert software, resulting in optimal experimental biomethane and bio-SA production of 85.7 % and 0.65 g/L, respectively, at a microwave dose of 1.45 W/g, Chaetomorpha ratio of 0.9 and pH value of 7.8. The study provides valuable insights into microwave control for promoting simultaneous methane enrichment and bio-SA production, potentially reducing costs associated with CO2 capture and storage and biogas upgrading.

Matched MeSH terms: Coculture Techniques
Fulltext Coculturing of Mucor plumbeus and Bacillus subtilis bacterium as an efficient fermentation strategy to enhance fungal lipid and gamma-linolenic acid (GLA) production

Mohamed H, Awad MF, Shah AM, Sadaqat B, Nazir Y, Naz T, et al.

Sci Rep, 2022 Jul 30;12(1):13111.
PMID: 35908106 DOI: 10.1038/s41598-022-17442-2

This study aimed to improve lipid and gamma-linolenic acid (GLA) production of an oleaginous fungus, Mucor plumbeus, through coculturing with Bacillus subtilis bacteria, optimising the environmental and nutritional culture conditions, and scaling them for batch fermentation. The maximum levels of biomass, lipid, fatty acid, and GLA in a 5 L bioreactor containing cellobiose and ammonium sulfate as the optimal carbon and nitrogen sources, respectively, achieved during the coculturing processes were 14.5 ± 0.4 g/L, 41.5 ± 1.3, 24 ± 0.8, and 20 ± 0.5%, respectively. This strategy uses cellobiose in place of glucose, decreasing production costs. The nutritional and abiotic factor results suggest that the highest production efficiency is achieved at 6.5 pH, 30 °C temperature, 10% (v/v) inoculum composition, 200 rpm agitation speed, and a 5-day incubation period. Interestingly, the GLA concentration of cocultures (20.0 ± 0.5%) was twofold higher than that of monocultures (8.27 ± 0.11%). More importantly, the GC chromatograms of cocultures indicated the presence of one additional peak corresponding to decanoic acid (5.32 ± 0.20%) that is absent in monocultures, indicating activation of silent gene clusters via cocultivation with bacteria. This study is the first to show that coculturing of Mucor plumbeus with Bacillus subtilis is a promising strategy with industrialisation potential for the production of GLA-rich microbial lipids and prospective biosynthesis of new products.

Matched MeSH terms: Coculture Techniques
Time-Lapse 2D Imaging of Phagocytic Activity in M1 Macrophage-4T1 Mouse Mammary Carcinoma Cells in Co-cultures

Zaidi NE, Shazali NAH, Chor ALT, Osman MA, Ibrahim K, Jaoi-Edward M, et al.

J Vis Exp, 2019 12 14.
PMID: 31885381 DOI: 10.3791/60281

Tumor-associated macrophages (TAMs) have been identified as an important component for tumor growth, invasion, metastasis, and resistance to cancer therapies. However, tumor-associated macrophages can be harmful to the tumor depending on the tumor microenvironment and can reversibly alter their phenotypic characteristics by either antagonizing the cytotoxic activity of immune cells or enhancing anti-tumor response. The molecular actions of macrophages and their interactions with tumor cells (e.g., phagocytosis) have not been extensively studied. Therefore, the interaction between immune cells (M1/M2-subtype TAM) and cancer cells in the tumor microenvironment is now a focus of cancer immunotherapy research. In the present study, a live cell coculture model of induced M1 macrophages and mouse mammary 4T1 carcinoma cells was developed to assess the phagocytic activity of macrophages using a time-lapse video feature using phase-contrast, fluorescent, and differential interference contrast (DIC) microscopy. The present method can observe and document multipoint live-cell imaging of phagocytosis. Phagocytosis of 4T1 cells by M1 macrophages can be observed using fluorescent microscopy before staining 4T1 cells with carboxyfluorescein succinimidyl ester (CFSE). The current publication describes how to coculture macrophages and tumor cells in a single imaging dish, polarize M1 macrophages, and record multipoint events of macrophages engulfing 4T1 cells during 13 h of coculture.

Matched MeSH terms: Coculture Techniques/methods*
Chondrogenesis of human adipose derived stem cells for future microtia repair using co-culture technique

Goh BS, Che Omar SN, Ubaidah MA, Saim L, Sulaiman S, Chua KH

Acta Otolaryngol, 2017 Apr;137(4):432-441.
PMID: 27900891 DOI: 10.1080/00016489.2016.1257151

CONCLUSION: In conclusion, these result showed HADSCs could differentiate into chondrocytes-like cells, dependent on signaling induced by TGF-β3 and chondrocytes. This is a promising result and showed that HADSCs is a potential source for future microtia repair. The technique of co-culture is a positive way forward to assist the microtia tissue.
OBJECTIVE: Reconstructive surgery for the repair of microtia still remains the greatest challenge among the surgeons. Its repair is associated with donor-site morbidity and the degree of infection is inevitable when using alloplastic prosthesis with uncertain long-term durability. Thus, human adipose derived stem cells (HADSCs) can be an alternative cell source for cartilage regeneration. This study aims to evaluate the chondrogenic potential of HADSCs cultured with transforming growth factor-beta (TGF-β) and interaction of auricular chondrocytes with HADSCs for new cartilage generation.
METHODS: Multi-lineages differentiation features of HADSCs were monitored by Alcian Blue, Alizarin Red, and Oil Red O staining for chondrogenic, adipogenic, and osteogenic differentiation capacity, respectively. Further, HADSCs alone were culture in medium added with TGF-β3; and human auricular chondrocytes were interacted indirectly in the culture with and without TGF-βs for up to 21 days, respectively. Cell morphology and chondrogenesis were monitored by inverted microscope. For cell viability, Alamar Blue assay was used to measure the cell viability and the changes in gene expression of auricular chondrocyte markers were determined by real-time polymerase chain reaction analysis. For the induction of chondrogenic differentiation, HADSCs showed a feature of aggregation and formed a dense matrix of proteoglycans. Staining results from Alizirin Red and Oil Red O indicated the HADSCs also successfully differentiated into adipogenic and osteogenic lineages after 21 days.
RESULTS: According to a previous study, HADSCs were strongly positive for the mesenchymal markers CD90, CD73, CD44, CD9, and histocompatibility antigen. The results showed HADSCs test groups (cultured with TGF-β3) displayed chondrocytes-like cells morphology with typical lacunae structure compared to the control group without TGF-β3 after 2 weeks. Additionally, the HADSCs test groups increased in cell viability; an increase in expression of chondrocytes-specific genes (collagen type II, aggrecan core protein, SOX 9 and elastin) compared to the control. This study found that human auricular chondrocytes cells and growth factor had a positive influence in inducing HADSCs chondrogenic effects, in terms of chondrogenic differentiate of feature, increase of cell viability, and up-regulated expression of chondrogenic genes.

Matched MeSH terms: Coculture Techniques*
Fulltext Development of Transgenic Papaya through Agrobacterium-Mediated Transformation

Azad MA, Rabbani MG, Amin L, Sidik NM

Int J Genomics, 2013;2013:235487.
PMID: 24066284 DOI: 10.1155/2013/235487

Transgenic papaya plants were regenerated from hypocotyls and immature zygotic embryo after cocultivation with Agrobacterium tumefaciens LBA-4404 carrying a binary plasmid vector system containing neomycin phosphotransferase (nptII) gene as the selectable marker and β-glucuronidase (GUS) as the reporter gene. The explants were co-cultivated with Agrobacterium tumefaciens on regeneration medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime for one week. The cocultivated explants were transferred into the final selection medium containing 500 mg/L carbenicillin + 200 mg/L cefotaxime + 50 mg/L kanamycin for callus induction as well as plant regeneration. The callus derived from the hypocotyls of Carica papaya cv. Shahi showed the highest positive GUS activities compared to Carica papaya cv. Ranchi. The transformed callus grew vigorously and formed embryos followed by transgenic plantlets successfully. The result of this study showed that the hypocotyls of C. papaya cv. Shahi and C. papaya cv. Ranchi are better explants for genetic transformation compared to immature embryos. The transformed C. papaya cv. Shahi also showed the maximum number of plant regeneration compared to that of C. papaya cv. Ranchi.

Matched MeSH terms: Coculture Techniques
Isolation of AHL-degrading bacteria from micro-algal cultures and their impact on algal growth and on virulence of Vibrio campbellii to prawn larvae

Pande GS, Natrah FM, Flandez AV, Kumar U, Niu Y, Bossier P, et al.

Appl Microbiol Biotechnol, 2015 Dec;99(24):10805-13.
PMID: 26344339 DOI: 10.1007/s00253-015-6918-1

Inactivation of quorum sensing (QS) signal molecules, such as acylhomoserine lactones (AHLs) of pathogenic bacteria, has been proposed as a novel method to combat bacterial diseases in aquaculture. Despite the importance of micro-algae for aquaculture, AHL degradation by bacteria associated with micro-algal cultures has thus far not been investigated. In this study, we isolated Pseudomonas sp. NFMI-T and Bacillus sp. NFMI-C from open cultures of the micro-algae Tetraselmis suecica and Chaetoceros muelleri, respectively. An AHL degradation assay showed that either monocultures or co-cultures of the isolates were able to degrade the AHL N-hexanoyl-L-homoserine lactone. In contrast, only Bacillus sp. NFMI-C was able to inactivate N-hydroxybutanoyl-L-homoserine lactone, the AHL produced by Vibrio campbellii. The isolated bacteria were able to persist for up to 3 weeks in conventionalized micro-algal cultures, indicating that they were able to establish and maintain themselves within open algal cultures. Using gnotobiotic algal cultures, we found that the isolates did not affect growth of the micro-algae from which they were isolated, whereas a mixture of both isolates increased the growth of Tetraselmis and decreased the growth of Chaetoceros. Finally, addition of Bacillus sp. NFMI-C to the rearing water of giant river prawn (Macrobrachium rosenbergii) larvae significantly improved survival of the larvae when challenged with pathogenic V. campbellii, whereas it had no effect on larval growth.

Matched MeSH terms: Coculture Techniques
Fulltext Biological Interaction Between Human Gingival Fibroblasts and Vascular Endothelial Cells for Angiogenesis: A Co-culture Perspective

Um Min Allah N, Berahim Z, Ahmad A, Kannan TP

Tissue Eng Regen Med, 2017 Oct;14(5):495-505.
PMID: 30603504 DOI: 10.1007/s13770-017-0065-y

Advancement in cell culture protocols, multidisciplinary research approach, and the need of clinical implication to reconstruct damaged or diseased tissues has led to the establishment of three-dimensional (3D) test systems for regeneration and repair. Regenerative therapies, including dental tissue engineering, have been pursued as a new prospect to repair and rebuild the diseased/lost oral tissues. Interactions between the different cell types, growth factors, and extracellular matrix components involved in angiogenesis are vital in the mechanisms of new vessel formation for tissue regeneration. In vitro pre-vascularization is one of the leading scopes in the tissue-engineering field. Vascularization strategies that are associated with co-culture systems have proved that there is communication between different cell types with mutual beneficial effects in vascularization and tissue regeneration in two-dimensional or 3D cultures. Endothelial cells with different cell populations, including osteoblasts, smooth muscle cells, and fibroblasts in a co-culture have shown their ability to advocate pre-vascularization. In this review, a co-culture perspective of human gingival fibroblasts and vascular endothelial cells is discussed with the main focus on vascularization and future perspective of this model in regeneration and repair.

Matched MeSH terms: Coculture Techniques
Fulltext Polycaprolactone-Based Scaffolds Facilitates Osteogenic Differentiation of Human Adipose-Derived Stem Cells in a Co-Culture System

Rozila I, Azari P, Munirah S, Safwani WKZW, Pingguan-Murphy B, Chua KH

Polymers (Basel), 2021 Feb 17;13(4).
PMID: 33671175 DOI: 10.3390/polym13040597

(1) Background: Stem cells in combination with scaffolds and bioactive molecules have made significant contributions to the regeneration of damaged bone tissues. A co-culture system can be effective in enhancing the proliferation rate and osteogenic differentiation of the stem cells. Hence, the aim of this study was to investigate the osteogenic differentiation of human adipose derived stem cells when co-cultured with human osteoblasts and seeded on polycaprolactone (PCL):hydroxyapatite (HA) scaffold; (2) Methods: Human adipose-derived stem cells (ASC) and human osteoblasts (HOB) were seeded in three different ratios of 1:2, 1:2 and 2:1 in the PCL-HA scaffolds. The osteogenic differentiation ability was evaluated based on cell morphology, proliferation rate, alkaline phosphatase (ALP) activity, calcium deposition and osteogenic genes expression levels using quantitative RT-PCR; (3) Results: The co-cultured of ASC/HOB in ratio 2:1 seeded on the PCL-HA scaffolds showed the most positive osteogenic differentiation as compared to other groups, which resulted in higher ALP activity, calcium deposition and osteogenic genes expression, particularly Runx, ALP and BSP. These genes indicate that the co-cultured ASC/HOB seeded on PCL-HA was at the early stage of osteogenic development; (4) Conclusions: The combination of co-culture system (ASC/HOB) and PCL-HA scaffolds promote osteogenic differentiation and early bone formation.

Matched MeSH terms: Coculture Techniques
Gene expression changes in human bronchial epithelial cells (BEAS-2B) and human pulmonary alveolar epithelial cells (HPAEpiC) after interaction with Cladosporium sphaerospermum

Lo SG, Wong SF, Mak JW, Choo KK, Ng KP

Med Mycol, 2020 Apr 01;58(3):333-340.
PMID: 31309220 DOI: 10.1093/mmy/myz061

Cladosporium is one of the most abundant spore. Fungi of this genus can cause respiratory allergy and intrabronchial lesion. We studied the differential expression of host genes after the interaction of Cladosporium sphaerospermum conidia with Human Bronchial Epithelial Cells (BEAS-2B) and Human Pulmonary Alveolar Epithelial Cells (HPAEpiC). C. sphaerospermum conidia were harvested and co-cultured with BEAS-2B cells or HPAEpiC cells for 48 hours respectively. This culture duration was chosen as it was associated with high germination rate. RNA was extracted from two biological replicates per treatment. RNA of BEAS-2B cells was used to assess changes in gene expression using AffymetrixGeneChip® Human Transcriptome Array 2.0. After co-culture with Cladosporium spores, 68 individual genes were found differentially expressed (P ≤ 0.05) and up-regulated ≥ 1.5 folds while 75 genes were found differentially expressed at ≤ -1.5 folds compared with controls. Reverse transcription and qPCR were performed on the RNA collected from both BEAS-2B cells and HPAEpiC cells to validate the microarray results with 7 genes. Based on the findings, infected pulmonary epithelial cells exhibited an increase in cell death-related genes and genes associated with innate immunity.

Matched MeSH terms: Coculture Techniques
Fulltext Enhancing food waste biodegradation rate in a food waste biodigester with the synergistic action of hydrolase-producing Bacillus paralicheniformis GRA2 and Bacillus velezensis TAP5 co-culture inoculation

Roslan MAM, Jefri NQUA, Ramlee N, Rahman NAA, Chong NHH, Bunawan H, et al.

Saudi J Biol Sci, 2021 May;28(5):3001-3012.
PMID: 34012331 DOI: 10.1016/j.sjbs.2021.02.041

Food waste (FW) minimization at the source by using food waste biodigester (FWBs) has a vast potential to lower down the impact of increasing organic fraction in municipal solid waste generation. To this end, this research sought to check the performance of locally isolated hydrolase-producing bacteria (HPB) to improve food waste biodegradation rate. Two under-explored HPB identified as Bacillus paralicheniformis GRA2 and Bacillus velezensis TAP5 were able to produce maximum amylase, cellulase, protease and lipase activities, and demonstrated a significant hydrolase synergy in co-culture fermentation. In vitro biodegradation analysis of both autoclaved and non-autoclaved FW revealed that the HPB inoculation was effective to degrade total solids (>62%), protein (>19%), total fat (>51), total sugar (>86%), reducing sugar (>38%) and starch (>50%) after 8-day incubation. All co-culture treatments were recorded superior to the respective monocultures and the uninoculated control. The results of FW biodegradation using batch-biodigester trial indicated that the 1500 mL and 1000 mL inoculum size of HPB inoculant reached a plateau on the 4th day, with gross biodegradation percentage (GBP) of >85% as compared to control (66.4%). The 1000 mL inoculum was sufficient to achieve the maximum GBP (>90%) of FW after an 8-day biodigestion in a FWB.

Matched MeSH terms: Coculture Techniques
Proteomics Analysis Revealed that Crosstalk between Helicobacter pylori and Streptococcus mitis May Enhance Bacterial Survival and Reduces Carcinogenesis

Khosravi Y, Loke MF, Goh KL, Vadivelu J

Front Microbiol, 2016;7:1462.
PMID: 27695448

Helicobacter pylori is the dominant species of the human gastric microbiota and is present in the stomach of more than half of the human population worldwide. Colonization by H. pylori causes persistent inflammatory response and H. pylori-induced gastritis is the strongest singular risk factor for the development of gastric adenocarcinoma. However, only a small proportion of infected individuals develop malignancy. Besides H. pylori, other microbial species have also been shown to be related to gastritis. We previously reported that interspecies microbial interaction between H. pylori and S. mitis resulted in alteration of their metabolite profiles. In this study, we followed up by analyzing the changing protein profiles of H. pylori and S. mitis by LC/Q-TOF mass spectrometry to understand the different response of the two bacterial species in a multi-species micro-environment. Differentially-expressed proteins in mono- and co-cultures could be mapped into 18 biological pathways. The number of proteins involve in RNA degradation, nucleotide excision repair, mismatch repair, and lipopolysaccharide (LPS) biosynthesis were increased in co-cultured H. pylori. On the other hand, fewer proteins involve in citrate cycle, glycolysis/ gluconeogenesis, aminoacyl-tRNA biosynthesis, translation, metabolism, and cell signaling were detected in co-cultured H. pylori. This is consistent with our previous observation that in the presence of S. mitis, H. pylori was transformed to coccoid. Interestingly, phosphoglycerate kinase (PGK), a major enzyme used in glycolysis, was found in abundance in co-cultured S. mitis and this may have enhanced the survival of S. mitis in the multi-species microenvironment. On the other hand, thioredoxin (TrxA) and other redox-regulating enzymes of H. pylori were less abundant in co-culture possibly suggesting reduced oxidative stress. Oxidative stress plays an important role in tissue damage and carcinogenesis. Using the in vitro co-culture model, this study emphasized the possibility that pathogen-microbiota interaction may have a protective effect against H. pylori-associated carcinogenesis.

Matched MeSH terms: Coculture Techniques
Chondrogenesis of adipose-derived stem cells with chondrocytes in low serum towards clinical application

Adila A Hamid, Satish Vaarman Jeyabalan, Aleza Omar, Nik Zattil Hanan Mohd Yasin, Wong TL, Liau LL, et al.

Sains Malaysiana, 2018;47:2369-2379.

Currently, fetal bovine serum (FBS) have been widely use in culture media to promote human cell proliferation. However,
the usage of FBS for cell therapy in clinical application was associated with the risk of viral and prion transmission as
well as immune rejection. To provide an option for this risk, this study was conducted to determine the effect of adipose
derived stem cells (ADSCs) co-culture with chondrocyte in promoting cell proliferation and chondrogenesis toward
FBS free condition. ADSCs co-cultured with chondrocyte at the ratio of 1:1, 2:1 and 1:2 were tested. Cell morphology
changes, cell proliferation and gene expression level of stemness (Oct4, FGF-4, Nanog) and chondrogenic (Collagen
Type II, ACP) were assessed. The results showed ADSCs in all co-culture groups changed morphology from fibroblastic
spindle to polygonal shape which resembled chondrocytes. The morphological changes were accompanied with increased
expression of chondrogenic genes; denoted chondrogenesis process. While maintaining expression of stemness genes
indicated continuation of cell proliferation. From the three co-culture groups tested; ADSCs and chondrocytes (1:1 ratio)
have been shown to exert better effects in promoting cell proliferation and chondrogenesis. In conclusion, ADSCs could
replace FBS to grow sufficient number of chondrogenic cells to repair cartilage injury in the near future. Further in vivo
study should be performed to test the effectiveness of this co-culture technique in cartilage injury repair.

Matched MeSH terms: Coculture Techniques
Fulltext IL-4 enhances IL-10 production in Th1 cells: implications for Th1 and Th2 regulation

Mitchell RE, Hassan M, Burton BR, Britton G, Hill EV, Verhagen J, et al.

Sci Rep, 2017 Sep 12;7(1):11315.
PMID: 28900244 DOI: 10.1038/s41598-017-11803-y

IL-10 is an immunomodulatory cytokine with a critical role in limiting inflammation in immune-mediated pathologies. The mechanisms leading to IL-10 expression by CD4(+) T cells are being elucidated, with several cytokines implicated. We explored the effect of IL-4 on the natural phenomenon of IL-10 production by a chronically stimulated antigen-specific population of differentiated Th1 cells. In vitro, IL-4 blockade inhibited while addition of exogenous IL-4 to Th1 cultures enhanced IL-10 production. In the in vivo setting of peptide immunotherapy leading to a chronically stimulated Th1 phenotype, lack of IL-4Rα inhibited the induction of IL-10. Exploring the interplay of Th1 and Th2 cells through co-culture, Th2-derived IL-4 promoted IL-10 expression by Th1 cultures, reducing their pathogenicity in vivo. Co-culture led to upregulated c-Maf expression with no decrease in the proportion of T-bet(+) cells in these cultures. Addition of IL-4 also reduced the encephalitogenic capacity of Th1 cultures. These data demonstrate that IL-4 contributes to IL-10 production and that Th2 cells modulate Th1 cultures towards a self-regulatory phenotype, contributing to the cross-regulation of Th1 and Th2 cells. These findings are important in the context of Th1 driven diseases since they reveal how the Th1 phenotype and function can be modulated by IL-4.

Matched MeSH terms: Coculture Techniques
Pseudomonas aeruginosa behaviour in polymicrobial communities: The competitive and cooperative interactions conducting to the exacerbation of infections

Al-Wrafy FA, Alariqi R, Noman EA, Al-Gheethi AA, Mutahar M

Microbiol Res, 2023 Mar;268:127298.
PMID: 36610273 DOI: 10.1016/j.micres.2022.127298

Pseudomonas aeruginosa is mostly associated with persistent infections and antibiotic resistance as a result of several factors, biofilms one of them. Microorganisms within the polymicrobial biofilm (PMB) reveal various transcriptional profiles and affect each other which might influence their pathogenicity and antibiotic tolerance and subsequent worsening of the biofilm infection. P. aeruginosa within PMB exhibits various behaviours toward other microorganisms, which may enhance or repress the virulence of these microbes. Microbial neighbours, in turn, may affect P. aeruginosa's virulence either positively or negatively. Such interactions among microorganisms lead to emerging persistent and antibiotic-resistant infections. This review highlights the relationship between P. aeruginosa and its microbial neighbours within the PMB in an attempt to better understand the mechanisms of polymicrobial interaction and the correlation between increased exacerbations of infection and the P. aeruginosa-microbe interaction. Researching in the literature that was carried out in vitro either in co-cultures or in the models to simulate the environment at the site of infection suggested that the interplay between P. aeruginosa and other microorganisms is one main reason for the worsening of the infection and which in turn requires a treatment approach different from that followed with P. aeruginosa mono-infection.

Matched MeSH terms: Coculture Techniques
Vimentin protein is a factor for decreasing breast cancer cell proliferation co-culture with human bone marrow-derived mesenchymal stem cells pre-treated with thiazolidinedione solutions

Kwok LS, Yian SS, Ismael LQ, Bee YTG, Harn GL, Yin KB

Mol Biol Rep, 2024 Feb 21;51(1):317.
PMID: 38381204 DOI: 10.1007/s11033-024-09269-z

BACKGROUND: Our previous study investigated the levels of soluble growth factors in the conditioned media of bone marrow-derived mesenchymal stem cells (BMSCs) pre-treated with thiazolidinedione solutions. The present study aimed to investigate the complex intracellular proteins extracted from BMSCs pre-treated with pioglitazone and/or rosiglitazone using proteomics.
METHODS: The proliferative effect of the identified protein on MCF-7 cells that interacted non-adhesively with BMSCs pre-treated with pioglitazone and/or rosiglitazone was evaluated using cell culture inserts and conditioned media. The mRNA expression of proliferation and lipid accumulation markers was also evaluated in the interacted MCF-7 cells by reverse transcription-quantitative PCR. Finally, the correlation between the identified protein and fibroblast growth factor 4 (FGF-4) protein in the conditioned media of the pre-treated BMSCs was evaluated by ELISA.
RESULTS: The present study identified vimentin as the specific protein among the complex intracellular proteins that likely plays a role in MCF-7 cell proliferation when the breast cancer cells interacted non-adhesively with BMSCs pre-treated with a combination of pioglitazone and rosiglitazone. The inhibition of this protein promoted the proliferation of MCF-7 cells when the breast cancer cells interacted with pre-treated BMSCs. Gene expression analysis indicated that pre-treatment of BMSCs with a combination of pioglitazone and rosiglitazone decreased the mRNA expression of Ki67 and proliferating cell nuclear antigen in MCF-7 cells. The pre-treatment did not induce mRNA expression of PPARγ, which is a sign of lipid accumulation. The level of vimentin protein was also associated with the FGF-4 protein expression level in the conditioned media of the pre-treated BMSCs. Bioinformatics analysis revealed that vimentin regulated the expression of FGF-4 through its interaction with SRY-box 2 and POU class 5 homeobox 1.
CONCLUSIONS: The present study identified a novel intracellular protein that may represent the promising target in pre-treated BMSCs to decrease the proliferation of breast cancer MCF-7 cells for human health and wellness.

Matched MeSH terms: Coculture Techniques

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