Biotechnology Letters最新文献

{"title":"Neutralizing anti-diphtheria toxin scFv produced by phage display","authors":"Ehsan Khalili, Mostafa Lakzaei, Mahdi Aminian","doi":"10.1007/s10529-024-03476-1","DOIUrl":"https://doi.org/10.1007/s10529-024-03476-1","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Background</h3><p>Diphtheria can be prevented by vaccination, but some epidemics occur in several places, and diphtheria’s threat is considerable. Administration of diphtheria antitoxin (DAT) produced from hyperimmunized animals is the most common treatment. Recombinant human antibody fragments such as single-chain variable fragments (scFv) produced by phage display library may introduce an interesting approach to overcome the limitations of the traditional antibody therapy. In the present study, B cells of immunized volunteers were used to construct a human single-chain fragment (HuscFv) library.</p><h3 data-test=\"abstract-sub-heading\">Materials and methods</h3><p>The library was constructed with the maximum combination of heavy and light chains. As an antigen, Diphtheria toxoid (DTd) was used in four-round phage bio-panning to select phage clones that display DTd bound HuscFv from the library. After panning, individual scFv clones were selected. Clones that were able to detect DTd in an initial screening assay were transferred to <i>Escherichia coli</i> HB2151 to express the scFvs and purification was followed by Ni metal ion affinity chromatography. Toxin neutralization test was performed on Vero cells. The reactivity of the soluble scFv with diphtheria toxin were done and affinity calculation based on Beatty method was calculated.</p><h3 data-test=\"abstract-sub-heading\">Results</h3><p>The size of the constructed scFv library was calculated to be 1.3 × 10⁶ members. Following four rounds of selection, 40 antibody clones were isolated which showed positive reactivity with DTd in an ELISA assay. Five clones were able to neutralize DTd in Vero cell assay. These neutralizing clones were used for soluble expression and purification of scFv fragments. Some of these soluble scFv fragments show neutralizing activity ranging from 0.6 to 1.2 µg against twofold cytotoxic dose of diphtheria toxin. The affinity constant of the selected scFv antibody was determined almost 10⁷ M⁻¹.</p><h3 data-test=\"abstract-sub-heading\">Conclusion</h3><p>This study describes the prosperous construction and isolation of scFv from the immune library, which specifically neutralizes diphtheria toxin. The HuscFv produced in this study can be a potential candidate to substitute the animal antibody for treating diphtheria and detecting toxins.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"38 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140572882","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pilot-scale production of Bacillus subtilis MSCL 897 spore biomass and antifungal secondary metabolites in a low-cost medium","authors":"Emils Bolmanis, Oskars Grigs, Elina Didrihsone, Maris Senkovs, Vizma Nikolajeva","doi":"10.1007/s10529-024-03481-4","DOIUrl":"https://doi.org/10.1007/s10529-024-03481-4","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Objectives</h3><p><i>Bacillus subtilis</i> is a plant growth promoting bacterium (PGPB) that acts as a microbial fertilizer and biocontrol agent, providing benefits such as boosting crop productivity and improving nutrient content. It is able to produce secondary metabolites and endospores simultaneously, enhancing its ability to survive in unfavorable conditions and eliminate competing microorganisms. Optimizing cultivation methods to produce <i>B. subtilis</i> MSCL 897 spores on an industrial scale, requires a suitable medium, typically made from food industry by-products, and optimal temperature and pH levels to achieve high vegetative cell and spore densities with maximum productivity.</p><h3 data-test=\"abstract-sub-heading\">Results</h3><p>This research demonstrates successful pilot-scale (100 L bioreactor) production of a biocontrol agent <i>B. subtilis</i> with good spore yields (1.5 × 10⁹ spores mL⁻¹) and a high degree of sporulation (&gt;80%) using a low-cost cultivation medium. Culture samples showed excellent antifungal activity (1.6–2.3 cm) against several phytopathogenic fungi. An improved methodology for inoculum preparation was investigated to ensure an optimal seed culture state prior to inoculation, promoting process batch-to-batch repeatability. Increasing the molasses concentration in the medium and operating the process in fed-batch mode with additional molasses feed, did not improve the overall spore yield, hence, process operation in batch mode with 10 g molasses L⁻¹ is preferred. Results also showed that the product quality was not significantly impacted for up to 12 months of storage at room temperature.</p><h3 data-test=\"abstract-sub-heading\">Conclusion</h3><p>An economically-feasible process for <i>B. subtilis</i>-based biocontrol agent production was successfully developed at the pilot scale.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"36 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140572868","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epigenetic modifiers as inducer of bioactive secondary metabolites in fungi","authors":"Vishal Sharma, Shivali Panjgotra, Nisha Sharma, Vidushi Abrol, Umesh Goutam, Sundeep Jaglan","doi":"10.1007/s10529-024-03478-z","DOIUrl":"https://doi.org/10.1007/s10529-024-03478-z","url":null,"abstract":"<p>Scientists are making efforts to search for new metabolites as they are essential lead molecules for the drug discovery, much required due to the evolution of multi drug resistance and new diseases. Moreover, higher production of known drugs is required because of the ever growing population. Microorganisms offer a vast collection of chemically distinct compounds that exhibit various biological functions. They play a crucial role in safeguarding crops, agriculture, and combating several infectious ailments and cancer. Research on fungi have grabbed a lot of attention after the discovery of penicillin, most of the compounds produced by fungi under normal cultivation conditions are discovered and now rarely new compounds are discovered. Treatment of fungi with the epigenetic modifiers has been becoming very popular since the last few years to boost the discovery of new molecules and enhance the production of already known molecules. Epigenetic literally means above genetics that actually does not alter the genome but alter its expression by altering the state of chromatin from heterochromatin to euchromatin. Chromatin in heterochromatin state usually doesn’t express because it is closely packed by histones in this state. Epigenetic modifiers loosen the packing of chromatin by inhibiting DNA methylation and histone deacetylation and thus permit the expression of genes that usually remain dormant. This study delves into the possibility of utilizing epigenetic modifying agents to generate pharmacologically significant secondary metabolites from fungi.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"145 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-04-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140572869","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improvement of cell growth in green algae Chlamydomonas reinhardtii through co-cultivation with yeast Saccharomyces cerevisiae","authors":"Yukino Karitani, Ryosuke Yamada, Takuya Matsumoto, Hiroyasu Ogino","doi":"10.1007/s10529-024-03483-2","DOIUrl":"https://doi.org/10.1007/s10529-024-03483-2","url":null,"abstract":"<h3 data-test=\"abstract-sub-heading\">Purpose</h3><p>CO₂ fixation methods using green algae have attracted considerable attention because they can be applied for the fixation of dilute CO₂ in the atmosphere. However, green algae generally exhibit low CO₂ fixation efficiency under atmospheric conditions. Therefore, it is a challenge to improve the CO₂ fixation efficiency of green algae under atmospheric conditions. Co-cultivation of certain microalgae with heterotrophic microorganisms can increase the growth potential of microalgae under atmospheric conditions. The objective of this study was to determine the culture conditions under which the growth potential of green algae <i>Chlamydomonas reinhardtii</i> is enhanced by co-culturing with the yeast <i>Saccharomyces cerevisiae</i>, and to identify the cause of the enhanced growth potential.</p><h3 data-test=\"abstract-sub-heading\">Results</h3><p>When <i>C. reinhardtii</i> and <i>S. cerevisiae</i> were co-cultured with an initial green algae to yeast inoculum ratio of 1:3, the cell concentration of <i>C. reinhardtii</i> reached 133 × 10⁵ cells/mL on day 18 of culture, which was 1.5 times higher than that of the monoculture. Transcriptome analysis revealed that the expression levels of 363 green algae and 815 yeast genes were altered through co-cultivation. These included genes responsible for ammonium transport and CO₂ enrichment mechanism in green algae and the genes responsible for glycolysis and stress responses in yeast.</p><h3 data-test=\"abstract-sub-heading\">Conclusion</h3><p>We successfully increased <i>C. reinhardtii</i> growth potential by co-culturing it with <i>S. cerevisiae</i>. The main reasons for this are likely to be an increase in inorganic nitrogen available to green algae via yeast metabolism and an increase in energy available for green algae growth instead of CO₂ enrichment.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":"2014 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2024-04-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140572875","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation of delignifying bacteria and optimization of microbial pretreatment of biomass for bioenergy.","authors":"B Rabi Prasad, Suman Polaki, Radha Krushna Padhi","doi":"10.1007/s10529-023-03463-y","DOIUrl":"10.1007/s10529-023-03463-y","url":null,"abstract":"<p><p>Microbial pretreatment of lignocellulosic biomass holds significant promise for environmentally friendly biofuel production, offering an alternative to fossil fuels. This study focused on the isolation and characterization of two novel delignifying bacteria, GIET1 and GIET2, to enhance cellulose accessibility by lignin degradation. Molecular characterization confirmed their genetic identities, providing valuable microbial resources for biofuel production. Our results revealed distinct preferences for temperature, pH, and incubation period for the two bacteria. Bacillus haynesii exhibited optimal performance under moderate conditions and shorter incubation period, making it suitable for rice straw and sugarcane bagasse pretreatment. In contrast, Paenibacillus alvei thrived at higher temperatures and slightly alkaline pH, requiring a longer incubation period ideal for corn stalk pretreatment. These strain-specific requirements highlight the importance of tailoring pretreatment conditions to specific feedstocks. Structural, chemical, and morphological analyses demonstrated that microbial pretreatment reduced the amorphous lignin, increasing cellulose crystallinity and accessibility. These findings underscore the potential of microbial pretreatment to enhance biofuel production by modifying the lignocellulosic biomass. Such environmentally friendly bioconversion processes offer sustainable and cleaner energy solutions. Further research to optimize these methods for scalability and broader application is necessary in the pursuit for more efficient and greener biofuel production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"183-199"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139511854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An inducible CRISPRi circuit for tunable dynamic regulation of gene expression in Saccharopolyspora erythraea.","authors":"Xing Jiang, Xiang Ke, Xiwei Tian, Ju Chu","doi":"10.1007/s10529-023-03462-z","DOIUrl":"10.1007/s10529-023-03462-z","url":null,"abstract":"<p><p>Actinomyces are gram-positive bacteria known for their valuable secondary metabolites. Redirecting metabolic flux towards desired products in actinomycetes requires precise and dynamic regulation of gene expression. In this study, we integrated the CRISPR interference (CRISPRi) system with a cumate-inducible promoter to develop an inducible gene downregulation method in Saccharopolyspora erythraea, a prominent erythromycin-producing actinobacterium. The functionality of the cumate-inducible promoter was validated using the gusA gene as a reporter, and the successful inducible expression of the dCas9 gene was confirmed. The developed inducible CRISPRi strategy was then employed to downregulate the expression of target genes rppA in the wild-type strain NRRL2338 and sucC in the high erythromycin-producing strain E3. Through dynamic control of sucC expression, a significant enhancement in erythromycin production was achieved in strain E3. This study demonstrated the effectiveness of an inducible gene downregulation approach using CRISPRi and a cumate-inducible promoter, providing valuable insights for optimizing natural product production in actinomyces.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"161-172"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139566374","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of bilirubin via whole-cell transformation utilizing recombinant Corynebacterium glutamicum expressing a β-glucuronidase from Staphylococcus sp. RLH1.","authors":"Wei Zhou, Yanan Cui, Mengyun Chen, Qijun Gao, Kai Bao, Yongzhong Wang, Min Zhang","doi":"10.1007/s10529-024-03468-1","DOIUrl":"10.1007/s10529-024-03468-1","url":null,"abstract":"<p><p>Bilirubin, a key active ingredient of bezoars with extensive clinical applications in China, is produced through a chemical process. However, this method suffers from inefficiency and adverse environmental impacts. To address this challenge, we present a novel and efficient approach for bilirubin production via whole-cell transformation. In this study, we employed Corynebacterium glutamicum ATCC13032 to express a β-glucuronidase (StGUS), an enzyme from Staphylococcus sp. RLH1 that effectively hydrolyzes conjugated bilirubin to bilirubin. Following the optimization of the biotransformation conditions, a remarkable conversion rate of 79.7% in the generation of bilirubin was obtained at temperate 40 °C, pH 7.0, 1 mM Mg²⁺ and 6 mM antioxidant NaHSO₃ after 12 h. These findings hold significant potential for establishing an industrially viable platform for large-scale bilirubin production.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"223-233"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139680640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dynamic cultivation of human mesenchymal stem/stromal cells for the production of extracellular vesicles in a 3D bioreactor system.","authors":"Ciarra Almeria, René Weiss, Maike Keck, Viktoria Weber, Cornelia Kasper, Dominik Egger","doi":"10.1007/s10529-024-03465-4","DOIUrl":"10.1007/s10529-024-03465-4","url":null,"abstract":"<p><strong>Purpose: </strong>3D cell culture and hypoxia have been demonstrated to increase the therapeutic effects of mesenchymal stem/stromal cells (MSCs)-derived extracellular vesicles (EVs). In this study, a process for the production of MSC-EVs in a novel 3D bioreactor system under normoxic and hypoxic conditions was established and the resulting EVs were characterized.</p><p><strong>Methods: </strong>Human adipose-derived MSCs were seeded and cultured on a 3D membrane in the VITVO® bioreactor system for 7 days. Afterwards, MSC-EVs were isolated and characterized via fluorescence nanoparticle tracking analysis, flow cytometry with staining against annexin V (Anx5) as a marker for EVs exposing phosphatidylserine, as well as CD73 and CD90 as MSC surface markers.</p><p><strong>Results: </strong>Cultivation of MSC in the VITVO® bioreactor system demonstrated a higher concentration of MSC-EVs from the 3D bioreactor (9.1 × 10⁹ ± 1.5 × 10⁹ and 9.7 × 10⁹ ± 3.1 × 10⁹ particles/mL) compared to static 2D culture (4.2 × 10⁹ ± 7.5 × 10⁸ and 3.9 × 10⁹ ± 3.0 × 10⁸ particles/mL) under normoxic and hypoxic conditions, respectively. Also, the particle-to-protein ratio as a measure for the purity of EVs increased from 3.3 × 10⁷ ± 1.1 × 10⁷ particles/µg protein in 2D to 1.6 × 10⁸ ± 8.3 × 10⁶ particles/µg protein in 3D. Total MSC-EVs as well as CD73^-CD90⁺ MSC-EVs were elevated in 2D normoxic conditions. The EV concentration and size did not differ significantly between normoxic and hypoxic conditions.</p><p><strong>Conclusion: </strong>The production of MSC-EVs in a 3D bioreactor system under hypoxic conditions resulted in increased EV concentration and purity. This system could be especially useful in screening culture conditions for the production of 3D-derived MSC-EVs.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"279-293"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10902030/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139721476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Construction of a bacteriophage-derived vector with potential applications in targeted drug delivery and cell imaging.","authors":"Mehdi Sharifi, Ali Akbar Alizadeh, Maryam Hamzeh Mivehroud, Siavoush Dastmalchi","doi":"10.1007/s10529-023-03455-y","DOIUrl":"10.1007/s10529-023-03455-y","url":null,"abstract":"<p><p>There is a strong relationship between the dysregulation of epidermal growth factor receptor (EGFR) and the development of epithelial-derived cancers. Therefore, EGFR has usually been considered the desired target for gene therapy. Here, we propose an approach for targeting EGFR-expressing cells by phage particles capable of displaying EGF and GFP as tumor-targeting and reporting elements, respectively. For this purpose, the superfolder GFP-EGF (sfGFP-EGF) coding sequence was inserted at the N-terminus of the pIII gene in the pIT₂ phagemid. The capability of the constructed phage to recognize EGFR-overexpressing cells was monitored by fluorescence microscopy, fluorescence-activated cell sorting (FACS), and cell-based ELISA experiments. FACS analysis showed a significant shift in the mean fluorescence intensity (MFI) of the cells treated with phage displaying sfGFP-EGF compared to phage displaying only sfGFP. The binding of phage displaying sfGFP-EGF to A-431 cells, monitored by fluorescence microscopy, indicated the formation of the sfGFP-EGF-EGFR complex on the surface of the treated cells. Cell-based ELISA experiments showed that phages displaying either EGF or sfGFP-EGF can specifically bind EGFR-expressing cells. The vector constructed in the current study has the potential to be engineered for gene delivery purposes as well as cell-based imaging for tumor detection.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"147-159"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139110743","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Differentiation, maturation, and collection of THP-1-derived dendritic cells based on a PEG hydrogel culture platform.","authors":"Jaeho Choi, Chang Seok Ki","doi":"10.1007/s10529-023-03457-w","DOIUrl":"10.1007/s10529-023-03457-w","url":null,"abstract":"<p><strong>Purpose: </strong>Dendritic cell (DC) is a spearhead responsible for immune response and surrounded by extracellular matrix in three-dimensional (3D) tissue. Nevertheless, conventional DC culture has relied on suspension or two-dimensional (2D) tissue culture plate (TCP)-based culture system. This culture condition often fails to recapitulate the physiological behavior of DC in real tissue. In this work, the effect of culture condition on DC physiology was explored with varying 3D hydrogel property (i.e., degradability, adhesion, and stiffness). In particular, DC differentiation and maturation in 3D were evaluated comparing the conventional TCP-based culture condition.</p><p><strong>Method: </strong>THP-1 cells were encapsulated in poly(ethylene glycol) (PEG) hydrogel via thiol-ene photocrosslinking with non-degradable or proteolytically degradable peptide crosslinker. Hydrogel stiffness was manipulated by controlling the concentration of crosslinker. The metabolic activities and cytotoxicity of the encapsulated cells were measured by resazurin and Live/Dead assays, respectively. Cell harvesting was conducted via enzymatic degradation using α-chymotrypsin, and differentiation and maturation of the liberated DCs were evaluated by quantitative polymerase chain reaction and flow cytometry.</p><p><strong>Results: </strong>THP-1 cells well proliferated in the soft degradable hydrogel with a higher metabolic activity. However, the stiff matrix inhibited cell growth in 3D. The gene expression assay indicated that the 3D hydrogel condition was superior to 2D culture in terms of differentiation and maturation of DC. Interestingly, the stiffness of matrix was important factor in DC function. In the stiff hydrogel, the expression levels of differentiation and maturation markers were higher compared to the low stiffness hydrogel. The mature DCs caged in the hydrogel matrix were harvested after short enzymatic digestion of hydrogel and the liberated cells had over 90% viability. The flow cytometric result revealed that the proportion of CD80 + /CD86 + cells from the stiff hydrogel was relatively higher than cells either from 2D or soft hydrogel in 3D.</p><p><strong>Conclusion: </strong>The collected evidence indicated that the proteolytically degradable PEG hydrogel matrix promoted DC differentiation and maturation. In addition, the matrix stiffness control could manipulate the marker expressions of differentiation and maturation. Particularly, the mature DC was successfully collected from the hydrogel matrix. These results highlighted the PEG hydrogel-based DC culture might be a useful tool for potential DC-based immunotherapies.</p>","PeriodicalId":8929,"journal":{"name":"Biotechnology Letters","volume":" ","pages":"235-247"},"PeriodicalIF":2.7,"publicationDate":"2024-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10901936/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139477899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}