In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"An in vitro cellular model for measuring the impact of thermal stress on Florida reef sponges.","authors":"Megan Conkling, Tobin Hindle, Zhixiao Xie, Weibo Liu, Timothy Moore, Shirley A Pomponi","doi":"10.1007/s11626-025-01034-1","DOIUrl":"https://doi.org/10.1007/s11626-025-01034-1","url":null,"abstract":"<p><p>Coral reefs are threatened by recurrent mortality incidents in their native habitats brought on by natural and anthropogenic stressors. Elevated temperature has been indicated as a major causing factor. Although ongoing research is focused on corals, sponges are an important benthic organism on coral reefs and are often overlooked. An accurate and standardized method is needed to determine the environmental limits and thresholds of sponges commonly found on coral reefs. We established an in vitro sponge cell model and evaluated the effect of elevated temperatures on primary cell cultures of five common Florida reef sponges-Agelas clathrodes, Aplysina fulva, Cliona varians, Geodia neptuni, and Xestospongia muta. Analysis of the results revealed that the impact of increased temperatures had no significant effect at the cellular level, but there are changes at the molecular level. Shifts in the sponges' transcriptomic profiles induced by increased temperatures, trigger processes related to signal transduction, apoptosis, and cell repair pathways. Further elevation of temperature corresponding to local extremes activated the immune response and programmed cell death. The results of the present study are based on both cellular and molecular data obtained from the in vitro cell model which highlight the minimal response of all five species to thermal stress, providing an insight into the mechanisms involved in the adaptive process. Furthermore, they suggest a resilience of these sponges to the current thermal extremes, but a combination of factors could still lead to a loss of sponges on reefs. This study forms the basis for use of in vitro sponge cell models to evaluate other environmental parameters and stressors on additional sponge species.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144127497","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":"mTORC1 signalling and protein synthesis are elevated in response to amino acids in human myotubes obtained from young, old, and old trained men.","authors":"Stephanie D Gagnon, Jiani Qian, Vladimir Belhac, Neil R W Martin","doi":"10.1007/s11626-025-01041-2","DOIUrl":"https://doi.org/10.1007/s11626-025-01041-2","url":null,"abstract":"<p><p>Ageing and reduced levels of physical activity are associated with desensitisation of skeletal muscle to the anabolic effects of amino acids. In vitro studies have indicated that many properties of skeletal muscle tissue are retained in human myotubes, including metabolic alterations associated with exercise and disease. However, the interaction between ageing and physical activity on amino acid sensing and growth has not been explored in human myotubes in vitro. Muscle-derived cells were isolated from biopsies taken from eight young (Y: 23.4 ± 1.9 yr), six older (O: 72.5 ± 5.0 yr), and nine older exercise trained (OT: 71.0 ± 4.1 yr, n = 9) men, and myotube cultures were generated and investigated for growth parameters and amino acid induced changes in mTORC1 signalling and protein synthesis. Our results indicated that muscle cell fusion was similar between groups, but myotube diameter was lower in cultures derived from O individuals. Despite this, mTORC1 signalling, as indicated by immunoblots for phosphorylation of mTOR^Ser2448, rpS6^Ser235/236, and 4E-BP1^Thr37/46 increased to a similar extent in response to amino acid availability in Y, O, and OT myotubes. Furthermore, measures of protein synthesis using the SUnSET assay were increased similarly between groups after the addition of amino acids. These data suggest that skeletal muscle desensitisation to amino acids with ageing is not observed in myotubes cultured in vitro, which could be reflective of the healthy individuals tested in our study or point towards the importance of the muscle niche in the impairments in muscle metabolism in ageing.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144110573","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":"Reduced myogenic differentiation capacity of satellite cell-derived myoblasts in male ICR mice compared with male C57BL/6 and BALB/c mice.","authors":"Takahiro Suzuki, Yuriko Nishi, Taku Koyama, Minori Nakada, Rio Arimatsu, Yusuke Komiya, Aoi Ogawa, Rika Osaki, Takahiro Maeno, Ai Saiga Egusa, Mako Nakamura, Ryuichi Tatsumi, Koichi Ojima, Takanori Nishimura","doi":"10.1007/s11626-025-01035-0","DOIUrl":"https://doi.org/10.1007/s11626-025-01035-0","url":null,"abstract":"<p><p>Many strains of wild-type laboratory mice have been developed for studies in the life sciences, including skeletal muscle cell biology. Muscle regeneration capacity differs among wild-type mouse strains. However, few studies have focused on whether myogenic stem cells (satellite cells) are directly related to mouse strain-dependent myoregeneration gaps using in vitro culture models. In this study, we selected three major wild-type mouse strains, CD1 (outbred; Jcl:ICR [ICR]), C57BL/6NJcl (inbred; B6), and BALB/cAJcl (inbred; C), which are widely used in laboratory experiments. Initially, we compared myotube fusion capabilities using satellite cell-derived myoblasts. The results showed that cell cultures isolated from male ICR mice could not efficiently form myotubes owing to low expression levels of myogenic regulatory factors (e.g., MyoD, myogenin, myocyte enhancer factor [MEF] 2A, and MEF2C) compared with B6 and C mouse strains. Next, we compared the myofiber-type compositions of muscle tissues and cultured myotubes among male mice from each of the three strains. Although each muscle tissue used for satellite cell isolation similarly expressed fast-twitch myofiber markers in all mouse strains, male ICR-derived myoblasts formed abundant amounts of slow-type myotubes. By contrast, myotubes from male B6 and C mice expressed substantial levels of fast-twitch myofiber markers. We also performed a comparative experiment in female ICR, B6, and C mouse strains, similar to the male mouse experiments. The myogenic differentiation potencies of myoblasts and myofiber-type compositions of myotubes in female mouse strains were similar. Thus, male ICR-derived satellite cells (myoblasts) had low myogenic differentiation potential, which may be associated with the tendency slow-twitch myotube formation.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144093482","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":"Novel use of a - 20°C cryoprotectant yields high viability and improved aggregation of marine sponge cells.","authors":"Elizabeth Urban-Gedamke, Megan Conkling, Cynthia Goodman, Xu Han, Shirley A Pomponi","doi":"10.1007/s11626-024-00959-3","DOIUrl":"10.1007/s11626-024-00959-3","url":null,"abstract":"","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"511-514"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141874691","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":"Challenges in cellular agriculture: lessons from Pacific white shrimp, Litopenaeus vannamei.","authors":"Catherine J Walsh, Tracy A Sherwood, Andrea M Tarnecki, Nicole R Rhody, Kevan L Main, Jessica Restivo","doi":"10.1007/s11626-024-01011-0","DOIUrl":"10.1007/s11626-024-01011-0","url":null,"abstract":"<p><p>The overall goal of this research was to develop an embryonic stem cell (ESC) line from the Pacific white shrimp, Litopenaeus vannamei, to support production of cell-based cultivated seafood products towards meeting a growing global demand for sustainable seafood. It was hypothesized that characteristics of ESCs, such as high proliferation and pluripotency, would facilitate development of a continuous cell line that could be triggered to differentiate into a muscle cell phenotype. The targeted approach was based on collection of ESCs from fertilized shrimp eggs at the blastomere stage. Various media, supplements, growth factors, and plate coatings were tested to achieve growth of the shrimp ESCs. Although successful in early culture, this manuscript describes substantial challenges encountered as cultures grew over time. The cell cultures were initially dominated by shrimp as indicated by 18S rDNA community analysis, but after multiple passages, thraustochytrids, a common contaminant of invertebrate cell culture, became the predominant cell type. Presence of shrimp cells was confirmed through species-specific primers for the cytochrome C oxidase subunit 1 gene. Presence of thraustochytrids was also confirmed using species-specific primers, morphological features, growth properties, and acriflavine staining. Unsuccessful attempts to eradicate thraustochytrid contamination prevented shrimp cells from thriving. The future of shrimp cell culture depends on eliminating culture contaminants while encouraging growth of shrimp ESCs.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"525-547"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143023288","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 and characterisation of two epithelial-like cell lines from the gills of Chrysophrys auratus (Australasian snapper) and Oncorhynchus tshawytscha (Chinook salmon) and their use in aquatic toxicology.","authors":"Björn Böhmert, Gavril L W Chong, Kim Lo, Michael Algie, Damon Colbert, Melissa D Jordan, Gabriella Stuart, Lyn M Wise, Lucy E J Lee, Niels C Bols, Georgina C Dowd","doi":"10.1007/s11626-024-00941-z","DOIUrl":"10.1007/s11626-024-00941-z","url":null,"abstract":"<p><p>In vitro gill models are becoming increasingly important in aquatic toxicology, yet the fish gill invitrome is underrepresented, encompassing approximately 0.1% of extant species. Here, we describe the establishment and characterisation of two gill-derived, epithelial-like cell lines isolated from fish species of significant importance to New Zealand: Chrysophrys auratus (Australasian snapper) and Oncorhynchus tshawytscha (Chinook salmon). Designated CAgill1PFR (Chrysophrys auratus, gill 1, Plant & Food Research) and OTgill1PFR (Oncorhynchus tshawytscha, gill 1, Plant & Food Research), these cell lines have each been passaged greater than each 70 times over several years and are considered spontaneously immortalised. Both cell lines required serum for growth and exhibited differential responses to basal media formulations. CAgill1PFR was sensitive to low temperatures (4 °C) but replicated at high temperatures (30 °C), whereas OTgill1PFR was sensitive to high temperatures but remained viable at low temperatures, mirroring the natural environment of their host species. Immunostaining revealed expression of epithelial cell markers cytokeratin and E-cadherin, alongside positivity for the mesenchymal cell marker, vimentin. CAgill1PFR was more sensitive to the environmental toxin 3,4 dichloroaniline than OTgill1PFR through measurements of metabolic activity, membrane integrity, and lysosomal function. Furthermore, CAgill1PFR produced less CYP1A activity, indicative of ongoing biotransformation processes, in response to beta-naphthoflavone than OTgill1PFR. These cell lines expand the toolbox of resources and emphasise the need for species-specific aquatic toxicology research.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"548-560"},"PeriodicalIF":1.5,"publicationDate":"2025-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12246032/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141579559","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":"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":"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}