In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"Conditions for establishing fin primary cell cultures in a wide range of ray-finned fishes.","authors":"Adauto Lima Cardoso, Jordana Inácio Nascimento Oliveira, João Pedro Silva Climaco, Natália Bortholazzi Venturelli, Camila do Nascimento Moreira, Cesar Martins","doi":"10.1007/s11626-024-00963-7","DOIUrl":"10.1007/s11626-024-00963-7","url":null,"abstract":"<p><p>Ray-finned fishes (Actinopterygii) represent the most diverse vertebrate lineage that show extensive variations in physiology, ways of life, and adaptations to marine and freshwater environments, and several species have been established as biological research models. The in vitro culture of cells is fundamental for several fields of biological research, being an alternative for studies that use animals. Hundreds of fish cell lines have been established using specific methods for each cell type and species. Here is described a protocol which can be used commonly for obtaining cell cultures from the caudal fin of a wide range of ray-finned fishes including marine and freshwater species. Conditions for sample collection, microbial disinfection, tissue dissociation, plating and incubation, cryopreservation and thawing, and karyotyping are described in detail. Primary cell cultures were developed for 20 species grouped into 12 different orders. Eleven of these species have been cultivated in vitro for the first time. In the beginning, the fish cell cultures showed different capacities of proliferation among them; however throughout the passages, most cultures began to have a similar proliferation rate. Throughout the passages, it was noticed that cells similar to fibroblasts began to predominate. The great proliferative ability of these cultures reveals their potential to become cell lines. The culture of A. mexicanus, for example, has been proliferating for months and is already in its 65th passage. Moreover, these cell cultures showed conserved diploid chromosome numbers in comparison with in vivo descriptions which suggest these cultures have stable karyotypes. Therefore, these cultures have potential to be used in several fields, such as toxicology, cytogenetics, genomics, pathology, immunology, cellular agriculture, and conservation, and this method has the potential to be expanded to species not yet tested, as well as to other organs.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"561-570"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141901597","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":"Efficacy determination of a disinfectant against channel catfish virus by in vitro and in vivo methods.","authors":"Suja Aarattuthodi, Brian Bosworth, Ganesh Kumar, Anita Nalamalapu","doi":"10.1007/s11626-024-01003-0","DOIUrl":"10.1007/s11626-024-01003-0","url":null,"abstract":"<p><p>Channel catfish virus (CCV) poses a significant threat to catfish culture. Lack of effective vaccines and antiviral treatments necessitates effective disinfection strategies to mitigate its spread. In vitro trials indicated the virus to be inactivated at high temperatures, but was infectious at 40°C. This study evaluated the efficacy of a commercial disinfectant against CCV using both in vitro and in vivo approaches. In vitro experiments assessed the virucidal activity of the disinfectant against CCV in channel catfish ovary (CCO) cells, while in vivo trials evaluated its effectiveness in reducing viral transmission and mortality among channel and hybrid catfish fingerlings. Results indicated that the disinfectant was effective in inactivating the virus at the tested concentrations and improved the survival of fish exposed to the virus. This study provides critical insights into selecting appropriate disinfection protocols to enhance biosecurity in catfish hatchery settings and to mitigate CCV transmission.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"582-590"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142909534","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":"Detection of Tilapia parvovirus in farm-reared tilapia in India and its isolation using fish cell lines.","authors":"Allahbagash Badhusha, Sivaraj Mithra, Gani Taju, Venkatesan Rajkumar, Seepoo Abdul Majeed, Selvam Suryakodi, Lekshmi Haridas, Divya Haridas, Pramoda Kumar Sahoo, Jyotirmaya Mohanty, Anirban Paul, Snatashree Mohanty, Devika Pillai, Vattiringal Jayadradhan Rejish Kumar, Azeez Sait Sahul Hameed","doi":"10.1007/s11626-024-01012-z","DOIUrl":"10.1007/s11626-024-01012-z","url":null,"abstract":"<p><p>Tilapia parvovirus (TiPV) is an emerging viral pathogen and responsible for severe economic loss in tilapia culture production. Lethargic, cutaneous haemorrhages; ocular lesions; discolouration of gill and cloudy eye and exophthalmia are common symptoms of TiPV. The TiPV-suspected tilapia fish were collected from grow-out ponds situated in different parts of Tamil Nadu, India, and screened for TiPV by PCR. The results showed the presence of TiPV in disease-suspected fish which was further confirmed by PCR using different primer sets specific to different genomic regions of TiPV. Sequence analysis of 534 bp of genomic region of TiPV showed 100% similarity with the sequence of TiPV strain of Thailand and India. TiPV was found in different organs including eggs of infected fish and showed the possibility of systemic infection and vertical transmission. Snakehead kidney (CSK), snubnose pompano fin (SPF) and tilapia heart (TH) cell lines showed susceptibility to TiPV. The viral replication in cell lines was confirmed by PCR, TiPV-specific cytopathic effect of Cowdry A inclusion bodies with clear halo surrounding them and infectivity experiment. The disease was reproduced in normal fish by intramuscular route using viral inoculum from TiPV-infected fish or virus multiplied in susceptible cell lines to satisfy Koch's postulates.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"601-613"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023290","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":"A cell line derived from the black soldier fly, Hermetia illucens (Diptera: Stratiomyidae).","authors":"Stephen Saathoff, Cynthia L Goodman, Eric Haas, Ian Mettelmann, David Stanley","doi":"10.1007/s11626-024-00934-y","DOIUrl":"10.1007/s11626-024-00934-y","url":null,"abstract":"<p><p>Insect cell lines are effective tools used in industry and academia. For example, they are used in screening potential insecticides, in making certain proteins for biomedical applications, and in basic research into insect biology. So far, there are no cell lines derived from the black soldier fly, Hermetia illucens (BSF). This may become an issue because BSFs are employed in a range of industrial and household processes. BSFs are used in producing biodiesel, in developing cosmetics and skin creams, and in the production of some medicines and animal feeds. BSF larvae process waste streams from a variety of sources into food for some animals and are also used in household composting. Our BSF cell line, designated BCIRL-HiE0122021-SGS, was developed from eggs using the medium CLG#2 (50% L-15 + 50% EX-CELL 420, with 9% FBS and antibiotics), with many other media being tested. This cell line consists of attached cells with a variety of morphologies and its identity was authenticated using CO1 barcoding. A growth curve was generated and the resulting doubling time was 118 h. We quantified the fatty acid methyl esters (FAMES) and recorded the expected range of saturated, monounsaturated, and polyunsaturated FAMEs, with only trace levels of lauric acid being noted. The BSF cell line is available free of charge by request.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"506-510"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141456510","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":"Development and characterization of polyvinyl alcohol/gelatin/chitosan hydrogel for tissue engineering and wound healing applications using a fish cell line model.","authors":"Sivaraj Mithra, Ali Asna Jabeen, Vinay Kumar, Seepoo Abdul Majeed, Manickam Balu Balaji, Sugumar Vimal, Dawood Mubeen Sultana, Sakvai Mohammed Safiullah, Gani Taju, Azeez Sait Sahul Hameed","doi":"10.1007/s11626-024-00996-y","DOIUrl":"10.1007/s11626-024-00996-y","url":null,"abstract":"<p><p>Chitosan-based hydrogels have gained considerable attention in biomedical research due to their inherent biocompatibility, biodegradability, and non-toxicity. When combined with polyvinyl alcohol (PVA), the resulting hydrogels exhibit superior mechanical strength, elasticity, and swelling capacity, making them highly suitable for a range of applications, including wound healing, tissue engineering, and controlled drug delivery. In this study, we synthesized and characterized a novel PVA/gelatin/chitosan (PVA/G/C) hydrogel composite using a series of analytical techniques such as Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray analysis (EDAX). The morphological, structural, and compositional analyses confirmed the successful formation of a homogenous, porous network conducive to cell proliferation and nutrient diffusion. In this study, polyvinyl alcohol/gelatin/chitosan-based hydrogels were prepared to study the potential for micro-tissue formation and wound healing application using Danio rerio gill (DrG) and Danio rerio fin (DrF) cell lines, respectively. Overall, the findings indicated the potential use of PVA/G/C hydrogel films as wound dressings. The idea of creating physically cross-linked hydrogels of PVA and chitosan without using any harmful organic chemicals or solvents is the novelty of this work. This study highlights the versatility and potential of PVA/G/C hydrogels, not only as a promising material for wound healing and drug delivery but also as an effective scaffold for tissue engineering applications.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"571-581"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142818006","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":"Using cationic liposomes as carriers for long dsRNA to trigger an antiviral response in rainbow trout cell lines.","authors":"Shayne J Oberhoffner, Dominique E Daniels, Erin Cooper, Aizah Ijaz, Starla A Richardson, Stephanie J DeWitte-Orr","doi":"10.1007/s11626-024-01002-1","DOIUrl":"10.1007/s11626-024-01002-1","url":null,"abstract":"<p><p>Long dsRNA induces the expression of type I interferons (IFNs) and IFN-stimulated genes (ISGs) to establish an antiviral state. When induced prophylactically, this antiviral state can reduce the severity and mortality of viral infections. One of the limiting factors in delivering dsRNA in animal models is the lack of an effective carrier that protects the dsRNA from degradation in the extracellular space. In this study, commercially available cationic liposomes composed of stearylamine, L-α-phosphatidylcholine, and cholesterol were analyzed for their ability to encapsulate and deliver a 621-bp dsRNA sequence. This encapsulated dsRNA was delivered to two Oncorhynchus mykiss cell lines, RTG-2 and RTgill-W1, to activate the IFN pathway and reduce chum salmon reovirus (CSV) infection. EMSA analysis revealed that the liposomes effectively encapsulated 55 and 800 µg/mL doses of dsRNA, remained stable when stored at 4°C and - 20°C, and protected the encapsulated dsRNA from degradation by RNase III. Cell viability assays determined that liposomes loaded with dsRNA were highly cytotoxic after 24 h of exposure. A shorter exposure of 2 h resulted in reduced cytotoxicity and enhanced expression of the ISG Mx1 in both dsRNA alone and dsRNA-liposome-treated cells; however, the elevated Mx1 induction was not sufficient in the dsRNA-liposome treatment group to provide protection against viral infection. Meanwhile, the unencapsulated dsRNA significantly reduced the CSV titer and amount of syncytia formation. Thus, while dsRNA represents an important immune modulator in fish cells, this liposome formulation is too toxic for antiviral applications.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"591-600"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142948201","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":"The impact of beauvericin on rainbow trout intestinal epithelial cells at different temperatures and dosing methods.","authors":"Vivian R Dayeh, Anita Solhaug, Mark E Hamilton, Laura E Linton, Lucy E J Lee, Niels C Bols","doi":"10.1007/s11626-025-01014-5","DOIUrl":"10.1007/s11626-025-01014-5","url":null,"abstract":"<p><p>Mycotoxins in aquatic feeds and their effects on fish are becoming more important in aquaculture, as fishmeal and fish oil in feeds are being replaced with more sustainable plant protein. Here, we investigated the potential of the mycotoxin, beauvericin (BEA), to impact the rainbow trout (RT) intestine by using cultures of the epithelial cell line, RTgutGC. BEA was dosed in different ways and exposed at temperatures ranging from 4 to 26 °C before being evaluated for cell viability by the metabolic reduction of Alamar Blue, by the accumulation of Neutral Red (lysosomal activity), cytotoxicity (CellTox Green), and for wound healing. BEA induces cell death in RTgutGC cells. The lysosomes are the main target (Neutral Red assay is the most sensitive) while cytotoxicity and plasma membrane rupture (CellTox Green) occur at considerably higher concentrations. BEA caused a dose-dependent decline in Neutral Red reading at all tested temperatures but Alamar Blue readings did not decline at 4 °C. Under these conditions, BEA appears to impair only lysosomal activity. Wound healing was reduced at 4, 10, and 26 °C compared to 18 °C. Also BEA treatment, at non-cytotoxic concentrations, reduced wound healing, but the temperature had little influence on this. Different carrier vehicles (methanol, DMSO) and exposure methods (passive or active dispersal) for BEA exposure were also studied. Here, methanol and passive dispersal gave comparable results to exposure with DMSO and active dispersal. In contrast, when DMSO was dosed with passive dispersal, immediate cytotoxicity in combination with BEA was induced.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"614-626"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143122812","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":"Cell Painting of insect gut cells for exploration of molecular responses of insect epithelia to insecticides.","authors":"Franziska Annabelle Hecker, Bruno Leggio, Tim Koenig, Karsten Niehaus, Sven Geibel","doi":"10.1007/s11626-025-01028-z","DOIUrl":"10.1007/s11626-025-01028-z","url":null,"abstract":"<p><p>Cell Painting is a sophisticated high-content imaging technique that has been predominantly applied to mammalian cells. Recent advancements have extended its applicability to the first insect cell line, the ovarian cell line Sf9, revealing significant insights into similarities and differences in cellular responses between different taxonomic groups. This study explores the utility of Cell Painting in Helicoverpa zea gut-derived cells, specifically the RP-HzGUT-AW1 cell line, to assess the specifics of insect epithelial cells in response to chemical treatments. Upon adaptation of the analysis pipeline to accommodate their unique morphology and characteristics, our investigations revealed distinct responses of RP-HzGUT-AW1 cells compared to the ovarian insect cell line Sf9. Variations were obtained not only in the dose-response behavior to treatments but also in the overall detectability of specific modes of action. Specifically, processes that relate to osmoregulation and the formation of epithelial structures showed the most significant and distinct responses. This suggests that the specific morphological and physiological attributes of these gut-derived insect cells contribute to unique phenotypic profiles, which enables in-depth interpretation of drug efficacy and safety in these models.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"515-524"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143648372","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":"Advancing marine invertebrate cell line research: four key knowledge gaps.","authors":"Baruch Rinkevich, Shirley A Pomponi","doi":"10.1007/s11626-025-01029-y","DOIUrl":"10.1007/s11626-025-01029-y","url":null,"abstract":"<p><p>Although cell cultures from marine invertebrates have great potential as valuable tools in various scientific fields, nearly all attempts to culture these cells in vitro have consistently failed, and the reasons for this remain unclear. The ongoing failure to develop stable, long-term cell cultures from marine invertebrates, despite varied species and methods employed, highlights significant knowledge gaps in understanding their in vitro requirements. These gaps impede progress, underscoring the complexity of marine invertebrate cells and the need for innovative approaches to overcome challenges in the field. When reviewing recent literature on the key data deficiencies and challenges behind the failure to develop marine invertebrate cell cultures, we identified and discussed four major knowledge gaps: (1) optimizing culture media, (2) strategies to extend stemness of isolated cells, (3) using \"omics\" to enhance cell culture, and (4) selecting suitable cell types for in vitro cultures. Bridging these gaps is crucial for advancing marine invertebrate cell culture systems. Yet, given the current state-of-the-art, addressing these gaps and advancing the discipline necessitate comprehensive, integrated, and species- or cell-specific strategies, along with close collaboration among laboratories working on diverse species.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"493-505"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12245970/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735794","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":"Cryopreservation of biological materials: applications and economic perspectives.","authors":"Suja Aarattuthodi, David Kang, Sanjay Kumar Gupta, Paula Chen, Bethany Redel, Moureen Matuha, Haitham Mohammed, Amit Kumar Sinha","doi":"10.1007/s11626-025-01027-0","DOIUrl":"https://doi.org/10.1007/s11626-025-01027-0","url":null,"abstract":"<p><p>Cryopreservation is a transformative technology that allows for the long-term storage of biological materials by cooling them to extremely low temperatures at which metabolic and biochemical processes are effectively slowed or halted. Cryopreservation utilizes various techniques to minimize ice crystal formation and cellular damage during freezing and thawing processes. This technology has broad applications in the fields of medicine, agriculture, and conservation, spanning across stem cell research, reproductive and regenerative medicine, organ transplantation, and cell-based therapies, each with significant economic implications. While current techniques and their associated costs present certain challenges, ongoing research advancements related to cryoprotectants, cooling methods, and automation promise to enhance efficiency and accessibility, potentially broadening the technology's impact across various sectors. This review focuses on the applications of cryopreservation, research advancements, and economic implications, emphasizing the importance of continued research to overcome the current limitations.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144063542","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}