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Montbretia flowers as a source of bioactive crocins: Biotechnology tools and delivery systems 作为生物活性藏红花的来源:生物技术工具和输送系统
Q1 Immunology and Microbiology Pub Date : 2025-03-28 DOI: 10.1016/j.btre.2025.e00891
Lucía Morote , Cristian Martínez Martínez Fajardo , María Mondéjar López , Elena Moreno-Gimenez , Ángela Rubio-Moraga , Olivia Costantina Demurtas , Gianfranco Diretto , Enrique Niza , Verónica Aragonés , Alberto López Jiménez , José-Antonio Daròs , Oussama Ahrazem , Lourdes Gómez-Gómez
Crocins are potent antioxidants with significant therapeutic potential, exhibiting anti-inflammatory, anticancer, and antidepressant properties. The ornamental plant Crocos-mia x crocosmiiflora is widely cultivated for its aromatic and vibrant flowers. In this study, we identified unique crocins as the primary pigments responsible for the flower's coloration. These metabolites predominantly consist of molecules with eight glucose units, followed by crocins containing six and seven glucose units. To elucidate the mo-lecular mechanisms underlying crocin biosynthesis in C. x crocosmiiflora, tran-scriptomic analysis was performed to identify key carotenoid cleavage dioxygenase (CCD) genes. Using Crocus sativus CsCCD2L gene as a bait, we identified a CCD transcript from the transcriptome data. Phylogenetic analysis revealed that the identified CCD belongs to the CCD2 subfamily, and it was designated as CroCCD2. Functional characterization of CroCCD2 was carried out using bacterial expression systems and Nicotiana benthamiana plants with a virus-mediated expression system. These experiments demonstrated that CroCCD2 efficiently converts the precursor zeaxanthin into crocetin, a key intermediate in crocin biosynthesis. Furthermore, we investigated the bioactivity of crocins and discovered that their anti-inflammatory effects depend on their vehiculation within exosomes or liposomes. This suggests that the transport mechanism is critical for the biological activity of crocins. Our findings highlight the specialization of CCD subfamilies in monocots and dicots for crocin biosynthesis and provide evidence of the anti-inflammatory activity of exosome-transported crocins. This study establishes a foundation for further research into the metabolic network of crocins in C. x crocosmiiflora and suggests that the CroCCD2 gene could be introduced into other crop plants to produce these bioactive apocarotenoids.
藏红花素是有效的抗氧化剂,具有显著的治疗潜力,表现出抗炎、抗癌和抗抑郁的特性。观赏植物crocosmiiflora因其芳香和鲜艳的花朵而被广泛种植。在这项研究中,我们确定了独特的藏红花色素作为主要色素负责花的颜色。这些代谢物主要由含有8个葡萄糖单位的分子组成,其次是含有6个和7个葡萄糖单位的藏红花。为了阐明藏红花素生物合成的分子机制,通过转录组学分析鉴定了类胡萝卜素裂解双加氧酶(CCD)的关键基因。以藏红花CsCCD2L基因为诱饵,从转录组数据中鉴定出一个CCD转录本。系统发育分析表明,该CCD属CCD2亚家族,命名为CroCCD2。用细菌表达系统和用病毒介导的烟植物表达系统对CroCCD2进行功能表征。这些实验证明CroCCD2有效地将前体玉米黄质转化为藏红花素,这是藏红花素生物合成的关键中间体。此外,我们研究了藏红花素的生物活性,发现它们的抗炎作用依赖于它们在外泌体或脂质体中的运载作用。这表明运输机制对藏红花素的生物活性至关重要。我们的研究结果强调了CCD亚家族在单子体和双子体中对藏红花素生物合成的专门化,并为外泌体运输藏红花素的抗炎活性提供了证据。本研究为进一步研究藏红花属植物中藏红花素的代谢网络奠定了基础,并提示可以将CroCCD2基因引入其他作物植物中以产生这些具有生物活性的类藏红花素。
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引用次数: 0
Cleaner horizons: Exploring advanced technologies for pollution remediation 更清洁的视野:探索先进的污染修复技术
Q1 Immunology and Microbiology Pub Date : 2025-03-28 DOI: 10.1016/j.btre.2025.e00890
Khadija Malik, Ashja Iftikhar, Quratulain Maqsood, Muhammad Rizwan Tariq, Shinawar Waseem Ali
Soil pollution causes many harmful effects by its contaminants or pollutants, which are known as soil pollutants. They are causing serious problems in plants as well as in humans. By entering into plants, harmful chemicals become part of the food chain. When humans consume contaminated food, it has harmful effects on human health. Pollutants are making soil unfit for living. Many techniques are being used for the remediation of soil pollution. Some are traditional techniques; some are innovative and effective as emerging science and technology are going on. In this review, we have discussed some significant methods, their aspects, and how they are playing their role in the remediation. Biological methods such as living organisms, chemical, and genetic manipulation are modern techniques that are being used for soil pollution remediation. Genetic manipulations sometimes change the enzyme processes, which enhance the whole activity by changing some of the proteins of organisms related to enzymes. Pollution remediation can be done by the process of bio-augmentation, which uses different types of strains of microbes for treatment. As there is an increase in the formation of OH compounds, advanced oxidation technologies are being introduced to treat them. Trace metals and heavy metals are also a big problem for soil pollution, which can be treated by phytoremediation techniques that use many different strategies. Nanoparticles are also being used for the treatment of compounds like nitrates, manganese, arsenic, etc. This review will guide you through the different technologies for soil pollution remediation.
土壤污染因其所含的污染物或污染物而产生许多有害影响,这些污染物被称为土壤污染物。它们给植物和人类都带来了严重的问题。通过进入植物,有害化学物质成为食物链的一部分。当人类食用受污染的食物时,它会对人体健康产生有害影响。污染物使土壤不适宜生物生存。许多技术被用于修复土壤污染。有些是传统技术;随着新兴科学技术的发展,有些是创新和有效的。在本文中,我们讨论了一些重要的方法,它们的方面,以及它们如何在修复中发挥作用。生物方法,如活生物体、化学和遗传操作是用于土壤污染修复的现代技术。基因操作有时会改变酶的过程,通过改变生物体中与酶相关的一些蛋白质来增强酶的整体活性。污染的修复可以通过生物强化的过程来完成,它使用不同类型的微生物菌株进行处理。随着氢氧根化合物形成的增加,先进的氧化技术正在被引入来处理它们。微量金属和重金属也是土壤污染的一个大问题,植物修复技术可以使用许多不同的策略来处理土壤污染。纳米颗粒也被用于处理硝酸盐、锰、砷等化合物。这篇综述将引导你了解不同的土壤污染修复技术。
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引用次数: 0
Optimized expression of human interleukin-15 in Nicotiana benthamiana and in vitro assessment of its activity on human keratinocytes 人白细胞介素-15在烟叶中的优化表达及其对人角质形成细胞活性的体外评价
Q1 Immunology and Microbiology Pub Date : 2025-03-25 DOI: 10.1016/j.btre.2025.e00889
Chalatorn Charnsatabut , Pipob Suwanchaikasem , Kaewta Rattanapisit , Iksen Iksen , Varisa Pongrakhananon , Christine Joy I. Bulaon , Waranyoo Phoolcharoen
Human interleukin-15 (hIL-15) is a cytokine essential for immune modulation with therapeutic applications in cancer and chronic wound healing. Although hIL-15 is commercially available, large-scale production studies remain limited. With promising clinical trial results, demand for hIL-15 is expected to rise. Plant expression systems offer a sustainable, low-cost alternative for rapid biopharmaceutical production. In this study, we optimized hIL-15 expression in Nicotiana benthamiana and assessed its physicochemical properties and biological activity. We fused hIL-15 to the Fc domain of human IgG1 for efficient purification. Through optimization of the pre- and post-infiltration conditions, we achieved transient expression and recovery at 4 dpi, yielding 33.8 µg/g fresh weight. Peptide mapping confirmed 97 % overall sequence coverage of the primary structure. Treatment with plant-produced hIL-15-Fc effectively promoted human keratinocyte HaCaT cell proliferation and migration in vitro. These findings demonstrated the potential of plant-based platforms for producing therapeutic recombinant hIL-15 that support wound healing.
人白细胞介素-15 (hIL-15)是一种重要的细胞因子,在癌症和慢性伤口愈合的治疗应用免疫调节。虽然hIL-15已商品化,但大规模生产研究仍然有限。随着有希望的临床试验结果,预计对hIL-15的需求将上升。植物表达系统为快速生物制药生产提供了一种可持续、低成本的替代方案。在本研究中,我们优化了hIL-15在烟叶中的表达,并对其理化性质和生物活性进行了评价。我们将hIL-15融合到人IgG1的Fc结构域以进行高效纯化。通过对浸前和浸后条件的优化,在4 dpi条件下实现了瞬时表达和恢复,产量为33.8µg/g鲜重。肽图谱证实97%的总序列覆盖初级结构。植物源hIL-15-Fc处理能有效促进人角质形成细胞HaCaT细胞的体外增殖和迁移。这些发现证明了基于植物的平台在生产支持伤口愈合的治疗性重组hIL-15方面的潜力。
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引用次数: 0
Chemical-induced variations in callus regenerated Atropa acuminata plants: A study on Ethyl Methanesulfonate effects 化学诱导的刺蒺藜愈伤组织变异:甲磺酸乙酯效应的研究
Q1 Immunology and Microbiology Pub Date : 2025-03-21 DOI: 10.1016/j.btre.2025.e00888
Shabeer Ahmad Dar , Ishfaq Shafi Khan , Gousia Majeed , Sumira Tyub , Irshad Ahmad Nawchoo , Azra Nahaid Kamili
Atropa acuminata Royle ex Lindl. a member of the solanaceae family is a critically endangered medicinal herb indigenous to the Himalayan region of Kashmir and revered for its therapeutic and economic significance. Ethyl Methanesulfonate (EMS), a chemical mutagen, is commonly used to induce mutations for plant breeding and research purposes. In this study, callus cultures derived from leaf explants of A. acuminata were cultivated on Murashige and Skoog (MS) medium supplemented with Thidiazuron (0.1 mg/l) and treated with varying concentrations of EMS (0.1%, 1.0%) over defined time intervals. The impact of EMS concentrations and exposure durations, as well as their interactions, on the characteristics of regenerated callus were rigorously examined. The proliferation of shoots exhibited a diminishing trend with escalating EMS concentrations. Additionally, EMS elicited substantial alterations in the morphological and physiological attributes of the regenerated plants. Among the fifteen traits evaluated, mutant lines designated as M10, M11, and M6 displayed the most pronounced variations in morphological and physiological parameters compared to the control. Notably, the mutant line M10 exhibited significantly elevated shoot dry weight, root fresh weight, chlorophyll, and carotenoid contents demonstrating up to 98%, 31%, 348%, and 642% enhancements respectively as compared to the control. These mutant lines hold promise for further exploitation in augmenting physiological characteristics and enhancing the yield and quality of terpene glycosides in A. acuminata.
火龙果(Atropa acuminata Royle ex Lindl)。茄科的一员,是一种极度濒危的草药,原产于克什米尔的喜马拉雅地区,因其治疗和经济意义而受到尊敬。甲基磺酸乙酯(EMS)是一种化学诱变剂,通常用于植物育种和研究目的的诱变。在本研究中,将刺荆叶片外植体的愈伤组织培养于Murashige和Skoog (MS)培养基上,外加0.1 mg/l的噻脲(Thidiazuron),并在规定的时间间隔内用不同浓度的EMS(0.1%、1.0%)处理。研究了EMS浓度、暴露时间及其相互作用对愈伤组织再生特性的影响。随着EMS浓度的升高,新梢的增殖呈减少趋势。此外,EMS引起了再生植株形态和生理特性的实质性改变。在15个性状中,M10、M11和M6突变系在形态和生理参数上的变化最为显著。值得注意的是,突变系M10的茎干重、根鲜重、叶绿素和类胡萝卜素含量均显著高于对照,分别提高了98%、31%、348%和642%。这些突变品系在改善荆芥生理特性、提高萜类苷产量和品质方面有进一步开发的希望。
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引用次数: 0
Design of a GFP reporter for splicing analysis in mammalian cells 用于哺乳动物细胞剪接分析的GFP报告基因的设计
Q1 Immunology and Microbiology Pub Date : 2025-03-13 DOI: 10.1016/j.btre.2025.e00887
Arthur T. Menezes, Helder Y. Nagasse, Hilan R. M. Lopes, Patricia P. Coltri
Eukaryotic genes are formed by exons and introns. Pre-mRNA splicing promotes exon ligation and intron removal and is performed by a specialized macromolecular machinery named spliceosome, composed of five small ribonucleoprotein particles (snRNPs) and more than one hundred proteins. The activity of this complex is highly accurate due to the coordinated activity of its components. Altered splicing has been related to the development of several diseases, including neurodegenerative disorders, such as amyotrophic lateral sclerosis, and different types of cancer. Detailed understanding of splicing regulation in eukaryotic cells can be achieved using splicing reporter systems. We designed a reporter plasmid suitable for splicing analysis in cultured mammalian cells. Our reporter is based on GFP expression, and the splicing outcome can be easily visualized by fluorescence microscopy. We quantified splicing activity in two human cell lines, HEK-293T and MDA-MB-231, confirming its suitability for use in live cells in culture.
真核生物基因由外显子和内含子组成。Pre-mRNA剪接促进外显子连接和内含子去除,由一种称为剪接体的特殊大分子机制完成,剪接体由五个小核糖核蛋白颗粒(snRNPs)和一百多种蛋白质组成。这种配合物的活性是高度准确的,由于其组分的协调活动。剪接的改变与几种疾病的发展有关,包括神经退行性疾病,如肌萎缩性侧索硬化症和不同类型的癌症。利用剪接报告系统可以详细了解真核细胞中的剪接调节。我们设计了一种适合于哺乳动物细胞剪接分析的报告质粒。我们的报告是基于GFP的表达,拼接结果可以很容易地在荧光显微镜下看到。我们量化了两种人类细胞系HEK-293T和MDA-MB-231的剪接活性,证实了其在活细胞培养中的适用性。
{"title":"Design of a GFP reporter for splicing analysis in mammalian cells","authors":"Arthur T. Menezes,&nbsp;Helder Y. Nagasse,&nbsp;Hilan R. M. Lopes,&nbsp;Patricia P. Coltri","doi":"10.1016/j.btre.2025.e00887","DOIUrl":"10.1016/j.btre.2025.e00887","url":null,"abstract":"<div><div>Eukaryotic genes are formed by exons and introns. Pre-mRNA splicing promotes exon ligation and intron removal and is performed by a specialized macromolecular machinery named spliceosome, composed of five small ribonucleoprotein particles (snRNPs) and more than one hundred proteins. The activity of this complex is highly accurate due to the coordinated activity of its components. Altered splicing has been related to the development of several diseases, including neurodegenerative disorders, such as amyotrophic lateral sclerosis, and different types of cancer. Detailed understanding of splicing regulation in eukaryotic cells can be achieved using splicing reporter systems. We designed a reporter plasmid suitable for splicing analysis in cultured mammalian cells. Our reporter is based on GFP expression, and the splicing outcome can be easily visualized by fluorescence microscopy. We quantified splicing activity in two human cell lines, HEK-293T and MDA-MB-231, confirming its suitability for use in live cells in culture.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00887"},"PeriodicalIF":0.0,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684353","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Exploring the multifaceted roles of resuscitation-promoting factors in tuberculosis: Implications for diagnosis, vaccine development, and drug targeting 探讨复苏促进因素在结核病中的多方面作用:对诊断、疫苗开发和药物靶向的影响
Q1 Immunology and Microbiology Pub Date : 2025-03-13 DOI: 10.1016/j.btre.2025.e00886
Gamze Tanriver , Salman Ali Khan , Artur Góra , Novel N Chegou , Shima Mahmoudi
Tuberculosis (TB) remains a significant global health challenge, necessitating continued research efforts to unravel its complex pathogenesis and advance diagnostic, therapeutic, and preventive strategies. Resuscitation-promoting factors (Rpfs) are peptidoglycan-hydrolyzing enzymes that have gained attention due to their key roles in TB infection dynamics. This review aims to provide a comprehensive overview of Rpfs in TB, highlighting their immunological roles, diagnostic potential, and implications for vaccine development and drug targeting through both in silico and experimental approaches. Rpfs exhibit diverse roles in TB, influencing bacterial resuscitation from dormancy, and immune modulation through interactions with host immune cells, such as dendritic cells, and they are potential targets for novel diagnostic and therapeutic interventions. Their ability to stimulate immune responses, particularly interferon-γ production by T cells, underscores their potential as vaccine candidates against TB. Moreover, Rpfs represent promising targets for drug discovery, with inhibitors potentially disrupting bacterial resuscitation and growth.
结核病(TB)仍然是一个重大的全球健康挑战,需要持续的研究努力来揭示其复杂的发病机制,并推进诊断、治疗和预防策略。复苏促进因子(Rpfs)是肽聚糖水解酶,因其在结核感染动力学中的关键作用而受到关注。本综述旨在全面概述Rpfs在结核病中的作用,强调它们的免疫学作用、诊断潜力,以及通过计算机和实验方法对疫苗开发和药物靶向的影响。Rpfs在结核病中表现出多种作用,影响细菌从休眠中复苏,并通过与宿主免疫细胞(如树突状细胞)的相互作用进行免疫调节,它们是新型诊断和治疗干预措施的潜在靶点。它们刺激免疫反应的能力,特别是T细胞产生干扰素γ的能力,强调了它们作为结核病候选疫苗的潜力。此外,Rpfs代表了药物发现的有希望的靶点,抑制剂可能会破坏细菌的复苏和生长。
{"title":"Exploring the multifaceted roles of resuscitation-promoting factors in tuberculosis: Implications for diagnosis, vaccine development, and drug targeting","authors":"Gamze Tanriver ,&nbsp;Salman Ali Khan ,&nbsp;Artur Góra ,&nbsp;Novel N Chegou ,&nbsp;Shima Mahmoudi","doi":"10.1016/j.btre.2025.e00886","DOIUrl":"10.1016/j.btre.2025.e00886","url":null,"abstract":"<div><div>Tuberculosis (TB) remains a significant global health challenge, necessitating continued research efforts to unravel its complex pathogenesis and advance diagnostic, therapeutic, and preventive strategies. Resuscitation-promoting factors (Rpfs) are peptidoglycan-hydrolyzing enzymes that have gained attention due to their key roles in TB infection dynamics. This review aims to provide a comprehensive overview of Rpfs in TB, highlighting their immunological roles, diagnostic potential, and implications for vaccine development and drug targeting through both in silico and experimental approaches. Rpfs exhibit diverse roles in TB, influencing bacterial resuscitation from dormancy, and immune modulation through interactions with host immune cells, such as dendritic cells, and they are potential targets for novel diagnostic and therapeutic interventions. Their ability to stimulate immune responses, particularly interferon-γ production by T cells, underscores their potential as vaccine candidates against TB. Moreover, Rpfs represent promising targets for drug discovery, with inhibitors potentially disrupting bacterial resuscitation and growth.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00886"},"PeriodicalIF":0.0,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143684354","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Retraction notice to “Fertilization value of municipal sewage sludge for Eucalyptus camaldulensis plants” [Biotechnology Reports 13 (2017) 8–12] 关于“城市污水污泥对桉植物的施肥价值”的撤回通知[生物技术报告13 (2017)8-12]
Q1 Immunology and Microbiology Pub Date : 2025-02-27 DOI: 10.1016/j.btre.2025.e00884
Soudani Leila , Maatoug Mhamed , Heilmeier Hermann , Kharytonov Mykola , Wiche Oliver , Moschner Christin , Onyshchenkoc Elena , Bouchenafa Nadia
{"title":"Retraction notice to “Fertilization value of municipal sewage sludge for Eucalyptus camaldulensis plants” [Biotechnology Reports 13 (2017) 8–12]","authors":"Soudani Leila ,&nbsp;Maatoug Mhamed ,&nbsp;Heilmeier Hermann ,&nbsp;Kharytonov Mykola ,&nbsp;Wiche Oliver ,&nbsp;Moschner Christin ,&nbsp;Onyshchenkoc Elena ,&nbsp;Bouchenafa Nadia","doi":"10.1016/j.btre.2025.e00884","DOIUrl":"10.1016/j.btre.2025.e00884","url":null,"abstract":"","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"47 ","pages":"Article e00884"},"PeriodicalIF":0.0,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144911821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Xenogeneic-free platform for the isolation and scalable expansion of human bladder smooth muscle cells 用于分离和扩增人膀胱平滑肌细胞的无异种平台
Q1 Immunology and Microbiology Pub Date : 2025-02-11 DOI: 10.1016/j.btre.2025.e00878
Teresa Franchi-Mendes , Marília Silva , Maria Catarina Carreira , Ana Luísa Cartaxo , Paulo Vale , Panagiotis Karakaidos , Apostolos Klinakis , Ana Fernandes-Platzgummer , Cláudia L. da Silva

Introduction

Smooth muscle cells (SMC) play a crucial role in bladder tissue engineering strategies. Scalable, Good Manufacturing Practice (GMP)-compliant platforms are essential for producing clinically relevant cell numbers.

Materials & Methods

A gamma-irradiated human platelet lysate (HPL) supplement was used to develop a xeno(geneic)-free process for the isolation and scalable expansion of human bladder-derived SMC.

Results

Cells were isolated using an explant-based technique and expanded ex vivo, expressing typical SMC markers (α-SMA, desmin, caldesmon and SM22-α). Cell culture was successfully scaled-up using spinner flasks combined with plastic microcarriers, starting with a 2.8 × 103 cells/cm2 inoculum (i.e. 1 × 106 cells). Cell-microcarrier adhesion rates exceeded 80% within 24 hours with fold expansion ranging from 3.5 to 16.8 after 7 days, dependent on donor variability. Harvested cells retained their SMC phenotype.

Conclusions

This xeno-free, GMP compliant process enables scalable manufacturing of human bladder-derived SMC while preserving cell identity, potentially advancing clinical applications in bladder engineering.
平滑肌细胞在膀胱组织工程策略中起着至关重要的作用。可扩展的、符合良好生产规范(GMP)的平台对于生产临床相关细胞数量至关重要。材料,方法采用γ射线辐照的人血小板裂解液(HPL)补充物,建立无xeno(基因)分离和扩展人膀胱源性SMC的工艺。结果采用外植体技术分离细胞并在体外扩增,表达典型的SMC标志物(α-SMA、desmin、caldesmon和SM22-α)。使用旋转烧瓶结合塑料微载体成功扩大细胞培养规模,接种量为2.8 × 103个细胞/cm2(即1 × 106个细胞)。细胞微载体黏附率在24小时内超过80%,7天后的倍数从3.5到16.8不等,取决于供体的变异。收获的细胞保留其SMC表型。结论:这种无xeno、符合GMP的工艺可以在保持细胞特性的同时可扩展地制造人类膀胱来源的SMC,潜在地推进膀胱工程的临床应用。
{"title":"Xenogeneic-free platform for the isolation and scalable expansion of human bladder smooth muscle cells","authors":"Teresa Franchi-Mendes ,&nbsp;Marília Silva ,&nbsp;Maria Catarina Carreira ,&nbsp;Ana Luísa Cartaxo ,&nbsp;Paulo Vale ,&nbsp;Panagiotis Karakaidos ,&nbsp;Apostolos Klinakis ,&nbsp;Ana Fernandes-Platzgummer ,&nbsp;Cláudia L. da Silva","doi":"10.1016/j.btre.2025.e00878","DOIUrl":"10.1016/j.btre.2025.e00878","url":null,"abstract":"<div><h3>Introduction</h3><div>Smooth muscle cells (SMC) play a crucial role in bladder tissue engineering strategies. Scalable, Good Manufacturing Practice (GMP)-compliant platforms are essential for producing clinically relevant cell numbers.</div></div><div><h3>Materials &amp; Methods</h3><div>A gamma-irradiated human platelet lysate (HPL) supplement was used to develop a xeno(geneic)-free process for the isolation and scalable expansion of human bladder-derived SMC.</div></div><div><h3>Results</h3><div>Cells were isolated using an explant-based technique and expanded ex vivo, expressing typical SMC markers (α-SMA, desmin, caldesmon and SM22-α). Cell culture was successfully scaled-up using spinner flasks combined with plastic microcarriers, starting with a 2.8 × 10<sup>3</sup> cells/cm<sup>2</sup> inoculum (i.e. 1 × 10<sup>6</sup> cells). Cell-microcarrier adhesion rates exceeded 80% within 24 hours with fold expansion ranging from 3.5 to 16.8 after 7 days, dependent on donor variability. Harvested cells retained their SMC phenotype.</div></div><div><h3>Conclusions</h3><div>This xeno-free, GMP compliant process enables scalable manufacturing of human bladder-derived SMC while preserving cell identity, potentially advancing clinical applications in bladder engineering.</div></div>","PeriodicalId":38117,"journal":{"name":"Biotechnology Reports","volume":"46 ","pages":"Article e00878"},"PeriodicalIF":0.0,"publicationDate":"2025-02-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143488299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Expanding black soldier fly (BSF; Hermetia illucens; Diptera: Stratiomyidae) in the developing world: Use of BSF larvae as a biological tool to recycle various organic biowastes for alternative protein production in Nepal 扩大黑兵蝇(BSF;Hermetia illucens;双翅目:层蛾科)在发展中国家:在尼泊尔,利用BSF幼虫作为一种生物工具,回收各种有机生物废物,用于替代蛋白质生产
Q1 Immunology and Microbiology Pub Date : 2025-02-10 DOI: 10.1016/j.btre.2025.e00879
Bhola Gautam , Sundar Tiwari , Min Raj Pokhrel , Jeffery K. Tomberlin , Prabhat Khanal
The growing global demand for food, particularly animal protein, is intensifying challenges related to food security and environmental sustainability. The increase in organic waste generation, coupled with inefficient waste management, is further deteriorating living conditions by negatively impacting the environment and public health, especially in developing nations. This study investigated the potential of black soldier fly larvae (BSFL) to recycle major daily organic waste fractions in Nepal. BSFL were exposed to seven different biowaste-based substrates locally sourced from fruit and vegetable markets, farms, and food industries. Additionally, the study evaluated rapeseed cake as a supplement to enhance BSFL growth and nutritional content. BSFL survival rates exceeded 80 % on food industry waste but dropped to 63 % on high-moisture substrates like vegetable waste. Mixed vegetable waste (14.7 mg/day) and bakery waste (11.5 mg/day) supported higher average daily weight gain, likely due to their better nutrient values (soluble carbohydrates and proteins). Although rapeseed cake alone hindered larval growth, its supplementation to biowastes improved growth, survival, and bioconversion rates, increased larval protein content up to 32 %, and reduced fat by 36 %. These findings indicate BSFL can effectively recycle diverse, locally available organic wastes in developing countries like Nepal, providing a sustainable source of domestic protein and contributing to feed security. As this is the first BSFL study in Nepal, further research is needed to elucidate the chemical and microbial safety of BSFL reared on biowastes and to develop technical solutions for commercial BSFL production in countries with a low-income economy.
全球对粮食,特别是动物蛋白的需求不断增长,加剧了与粮食安全和环境可持续性有关的挑战。有机废物产生的增加,加上废物管理效率低下,对环境和公众健康产生负面影响,进一步恶化了生活条件,特别是在发展中国家。本研究调查了黑兵蝇幼虫(BSFL)回收尼泊尔主要日常有机废物馏分的潜力。BSFL接触了七种不同的生物废物基基质,这些基质来自当地的水果和蔬菜市场、农场和食品工业。此外,研究还评估了油菜籽饼作为一种补充剂,可以促进BSFL的生长和营养含量。BSFL在食品工业废弃物上的存活率超过80%,但在蔬菜废弃物等高水分基质上的存活率降至63%。混合蔬菜废物(14.7毫克/天)和烘焙废物(11.5毫克/天)支持更高的平均每日体重增加,可能是因为它们具有更好的营养价值(可溶性碳水化合物和蛋白质)。虽然单独使用油菜籽饼会阻碍幼虫的生长,但在油菜籽饼中添加生物废物可以改善幼虫的生长、存活和生物转化率,使幼虫的蛋白质含量提高32%,脂肪含量降低36%。这些发现表明,在尼泊尔等发展中国家,BSFL可以有效地回收各种当地可用的有机废物,为国内蛋白质提供可持续来源,并有助于饲料安全。由于这是尼泊尔的第一个BSFL研究,需要进一步的研究来阐明利用生物废物饲养的BSFL的化学和微生物安全性,并为低收入国家的BSFL商业化生产开发技术解决方案。
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引用次数: 0
Microalgae strains isolated from piggery wastewater in Ecuador: Effective nitrogen compound removal and growth potential in extremophile conditions 从厄瓜多尔养猪场废水中分离的微藻菌株:在极端微生物条件下有效去除氮化合物和生长潜力
Q1 Immunology and Microbiology Pub Date : 2025-02-10 DOI: 10.1016/j.btre.2025.e00883
Karla Flores-Zambrano , Wilson Tapia , Pablo Castillejo
Effluents generated by anthropogenic activities are a significant source of pollution and eutrophication in natural water bodies. In Ecuador, the increase in pig production has exacerbated this issue due to the untreated discharge of pig effluents. This study focused on the characterization of native microalgae present in pig effluents and the evaluation of their capacity to remove nitrogenous compounds under various conditions, with the aim of identifying efficient strains for phycoremediation. Four microalgal strains were isolated and molecularly identified as Radiococcus polycoccus, Chlorolobion braunii, Micractinium sp., and Desmodesmus multivariabilis. The cultures were exposed to initial concentrations of 100 mg L-1 N-NH₄ and 49.97 mg L-1 N-NO₃ for 12 days to assess their cellular growth and nutrient removal rates. Growth kinetics were analyzed under conditions of 2000 mg L-1 N-NH₄ and extreme pH levels of 3 and 10. Chlorolobion braunii demonstrated the highest productivity, achieving a removal of 67.73 % of N-NH₄ and 30.59 % of N-NO₃, and reached the highest cellular density under extreme ammonium conditions, being the only strain capable of growing at acidic pH. Conversely, Micractinium sp. exhibited the highest growth under alkaline conditions. These results highlight the promising potential of native microalgae from pig effluents for wastewater remediation and their adaptation to environmental conditions.
人为活动产生的污水是自然水体污染和富营养化的重要来源。在厄瓜多尔,由于未经处理的猪粪排放,生猪产量的增加加剧了这一问题。本研究重点研究了猪废水中天然微藻的特性,并评估了它们在不同条件下去除含氮化合物的能力,目的是确定有效的藻修复菌株。分离得到4株微藻,经分子鉴定分别为多球菌(Radiococcus polycoccus)、褐绿菌(Chlorolobion braunii)、微藻(micractiinium sp.)和多变桥胞菌(desmodesabilis)。将培养物暴露在初始浓度为100 mg L-1 N-NH₄和49.97 mg L-1 N-NO₃的培养基中12天,以评估它们的细胞生长和营养去除率。在2000 mg L-1 N-NH _4和极端pH值3和10条件下,分析了生长动力学。cloolobion braunii的产率最高,能脱除67.73%的N-NH₄和30.59%的N-NO₃,并且在极端铵态条件下达到最高的细胞密度,是唯一能在酸性条件下生长的菌株。相反,Micractinium sp.在碱性条件下生长最快。这些结果突出了猪废水中原生微藻在废水修复和环境条件适应方面的巨大潜力。
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引用次数: 0
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