In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"Establishment and characterization of a continuous cell line from eye of the brown-marbled grouper (Epinephelus fuscoguttatus) for virological studies.","authors":"Jiqing Zheng, Jinwu Wang, Ting Xue","doi":"10.1007/s11626-025-01140-0","DOIUrl":"10.1007/s11626-025-01140-0","url":null,"abstract":"<p><p>Fish cell lines are indispensable tools for virology, biotechnology, and toxicology research. This study established a new marine fish cell line, designated EfE, from the eye tissue of the brown-marbled grouper (Epinephelus fuscoguttatus). The EfE cell line has been stably subcultured for over 70 passages in vitro for more than 300 days. It proliferated optimally in Leibovitz's L-15 medium supplemented with 15% fetal bovine serum at 28°C. Species origin was confirmed by molecular analysis of the mitochondrial CO1 gene. Chromosome analysis revealed a diploid count of 48, which is consistent with the karyotype of E. fuscoguttatus. The cell line demonstrated high transfection efficiency (25.6%) with a pEGFP-N1 plasmid, indicating its potential for genetic manipulation. In virus susceptibility tests, EfE cells were highly permissive to red-spotted grouper nervous necrosis virus (RGNNV), developing severe cytopathic effects (CPE), including extensive vacuolation, cell rounding, and detachment. Viral replication was further confirmed by semi-quantitative RT-PCR and the observation of virus particles via transmission electron microscopy (TEM). In conclusion, the novel EfE cell line provides a valuable in vitro model for virus isolation, propagation, investigation of pathogenic mechanisms, and genetic studies.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"898-908"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145989111","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":"In vitro culture for long-term maintenance of embryonic insect cells from the house cricket Acheta domesticus.","authors":"Stephen A Schwartz, Sophia M Letcher, Noah Novick, Shwe Yee Pwint, David L Kaplan","doi":"10.1007/s11626-026-01174-y","DOIUrl":"10.1007/s11626-026-01174-y","url":null,"abstract":"<p><p>Insect cell culture has become foundational for many areas of research and biotechnological innovation, yet major taxonomic gaps persist, particularly among orthopteran species. Here, we report the isolation and long-term maintenance of primary embryonic cells derived from the house cricket, Acheta domesticus. By isolating cells from late-stage embryos and culturing them at 29°C in Shields and Sang M3 insect cell media supplemented with 1 mg/mL yeast extract, 2.5 mg/mL Bacto Peptone, 100 µg/mL primocin, and 20% heat inactivated FBS, we were able to establish robust primary cultures of small, rounded cells. Passaging via mechanical sloughing preserved cell viability and supported 37.7 cumulative doublings over 287 days, thus, an average doubling time of 182.4 h. The results highlight the importance of conditioned media for sustaining proliferation, while also identifying challenges related to metabolite accumulation and slow doubling times. These preliminary findings provide an avenue for expanding the diversity of insect cell lines, with promising implications for both cellular agriculture and fundamental biotechnology research.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"952-959"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147591750","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":"Coating materials enhance urochordate primary cell culture adherence.","authors":"Andy Qarri, Dietmar Kültz, Alison M Gardell, Yiqun Su, Yuval Rinkevich, Baruch Rinkevich","doi":"10.1007/s11626-026-01213-8","DOIUrl":"10.1007/s11626-026-01213-8","url":null,"abstract":"<p><p>Marine invertebrate cell cultures are a potential source for diverse biotechnological applications, given the wide range of bioactive compounds they synthesize and accumulate. Yet, the number of established marine invertebrate cell culture systems remains limited compared with those of insects and vertebrates, particularly with respect to adherent cell cultures. Here we studied the in vitro adherence of circulating blood cells from the colonial ascidian Botryllus schlosseri. Two experimental approaches were employed, seeding blood cells either alone (setup 1) or in combination with tissue fragments (setup 2), using three basal media (DMEM, DMEM/F-12, RPMI) on culture plates coated with either Poly-L/D-lysine, gelatin, collagen, or laminin. Each experiment lasted for up to 3 d. Setup 1 results reveal that Botryllus cells remain viable and can adhere to coated surfaces in all tested media. Collagen- and laminin-coated plates supported longer-term cultures, whereas Poly-D (or L)-lysine coatings were more suitable for short-term studies. Among the basal media, RPMI and DMEM/F12 most effectively supported cell attachment. Setup 2 plates consistently showed higher cell adherence compared to setup 1, suggesting that tissue-derived factors may enhance attachment. Overall, circulating Botryllus cells demonstrate the capacity for substrate adhesion in vitro, offering a foundation for the development of adherent cell cultures.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"1013-1025"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148470254","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 characterisation of a novel cell line derived from coho salmon (Oncorhynchus kisutch).","authors":"Mari Austad, Jacob Seilø Torgersen, Beate Beatriz Furevik, Lucy E J Lee, Guro Katrine Sandvik","doi":"10.1007/s11626-025-01113-3","DOIUrl":"10.1007/s11626-025-01113-3","url":null,"abstract":"<p><p>Coho salmon (Oncorhynchus kisutch) is an important salmonid species differing from other salmonids in its tolerance and response to pathogens endemic to the aquaculture industry, such as infectious pancreatic necrosis virus (IPNV) and infectious salmon anaemia virus (ISAV). Consequently, coho salmon has become a subject of increased scientific interest to investigate the underlying genetic mechanisms behind these and other host-pathogen interactions. Currently, most research studying coho salmon has been conducted using live animal models as there have been few in vitro tools readily available. Here, we present the first cell line from an adult coho salmon, Coho Salmon Fibroblast-Like 1 Norway-Canada (CSFL-1NC) and its preliminary characterisation. CSFL-1NC is a homogenous, spontaneously immortalised cell line from the pectoral fin of a wild adult coho salmon, with a consistent and stable fibroblastic morphology. The cell line has a relatively stable transcriptome across several passages, with high expression of key fibroblastic marker genes, displays rapid migration, and can be genetically manipulated both by transfection and transduction with varying efficiency using plasmids, lentivirus, and/or CRISPR methodology. Virus challenges show clear susceptibility to IPNV as evidenced by cytopathic effects and efficient viral replication, yet it shows little to no response when exposed to ISAV (HPRD).</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"847-862"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13582294/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145285990","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":"Establishment and characterization of an ovarian cell line from red seabream (Pagrus major) and its application in fish toxicology.","authors":"Yucong Yang, Yitong Zhang, Mingyang Liu, Guixing Wang, Zhongwei He, Yufeng Liu, Heng Zhang, Tian Han, Yuqin Ren, Jilun Hou","doi":"10.1007/s11626-026-01208-5","DOIUrl":"10.1007/s11626-026-01208-5","url":null,"abstract":"<p><p>Fish cell lines serve as ideal in vitro models for investigating the toxicity of environmental pollutants in fish. In this study, a novel cell line (PmOVC) derived from the ovary of red seabream (Pagrus major) was successfully established. Using polystyrene microplastics (PS-MPs)-a typical environmental pollutant-as the test substance, the toxicity sensitivity of the PmOVC cell line was systematically assessed from the dimensions of particle size intake and cell activity. The results showed that the PmOVC cells exhibited robust growth in M199 medium supplemented with 15% FBS at 23°C and high exogenous gene transfection efficiency and showed high sensitivity to the toxic effects of microplastics. Specifically, the smaller the particle size of microplastics, the higher the internalization efficiency of cells, among which 100 nm PS-MPs could be completely endocytosed into the cytoplasm; the concentration of PS-MPs was significantly positively correlated with the apoptosis rate of cells (R² ≥ 0.8). In summary, the established PmOVC cell line provides a critical in vitro tool for in-depth investigation of the toxicological responses and underlying mechanisms of fish to environmental pollutants.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"1002-1012"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148360666","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":"Complete ectosome formation from the choanosome during whole-body regeneration of the sponge Hymeniacidon heliophila (Wilson 1911) using an in vitro microexplant model.","authors":"Cristiano C Coutinho, Fernando B M Henriques, Márcia Attias, Claudia Mermelstein, Manoel L Costa","doi":"10.1007/s11626-026-01152-4","DOIUrl":"10.1007/s11626-026-01152-4","url":null,"abstract":"<p><p>The regeneration of the sponge Hymeniacidon heliophila was investigated using the in vitro microexplant model. This model differs from cell dissociation, surface ablation, and fragment culture models because it preserves the choanosome attached to the substrate, maintaining the apical-basal axis while entirely lacking the ectosome and the water flow polarity (ostia-osculum). The complete ectosome regenerative development was assessed using stereoscopy and scanning electron microscopy until complete adult formation. The observed stages of ectosome regeneration from the preserved choanosome align with existing data on sponge regeneration. Epithelization begins ubiquitously across the microexplant surface and expands to fully encase the microexplant. Two key processes were noticed: (1) the formation of an extracellular layer at the surface and (2) the appearance of discrete patches of pinacoderm over the extracellular matrix, probably by mesenchymal-to-epithelial transition. In more advanced regeneration stages, ectosomal spicules align transversely to the pinacoderm, with their tips protruding outward. These spicules provide structural support for the developing subdermal space while the choanosome concurrently becomes functional. Additionally, outer cells were identified in both adult microexplants and naturally growing individuals. Further efforts are needed to distinguish whether these outer cells are epibionts or autologous. As final considerations, the data were discussed from an evolutionary perspective that regeneration in sponges is an inherent feature linked to the emergence of multicellularity in animals. This process represents a primordial mechanism for stabilizing metazoan multicellular organization.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"923-936"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13582140/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146257981","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":"Isolation of coelomocyte from sea urchin Echinometra mathaei: optimization of culture condition.","authors":"Fatemeh Piryaei, Pargol Ghavam Mostafavi, Razieh Dalirfardouei, Fahimeh Piryaei","doi":"10.1007/s11626-025-01114-2","DOIUrl":"10.1007/s11626-025-01114-2","url":null,"abstract":"<p><p>Rapid growth of the aquaculture industry is hampered by infectious diseases in marine invertebrates, causing economic losses. Marine invertebrate cell cultures offer tools to evaluate biological properties and cellular responses in different conditions. Long-term culture aims to isolate tissue-specific cells and identify bioactive compounds from stem cells. Echinometra mathaei, known as Persian Gulf sea urchin, has lots of benefits in various fields including aquaculture, embryology, and evolutionary biology. However, its cell culture faces challenges due to poorly characterized microenvironmental and specific cultivation requirements. This study aims to establish and optimize a long-term cell culture for coelomocyte derived from E. mathaei, focusing on the characterization of microenvironment conditions to overcome the limitations of current marine invertebrate cell culture. After the collection of E. mathaei from Lark Island, Persian Gulf, Iran, and their acclimatization in artificial seawater, coelomocytes were isolated from different sources including the coelomic fluid, the coelomic epithelium, and the axial organ. Various cell dissociation methods, culture media, growth supplements, culture dishes, and physical conditions were tested to determine optimal conditions for coelomocyte in vitro culture. Moreover, coelomocytes were differentiated to pigment-producing cells, and naphthoquinone pigments were extracted and identified using spectrophotometry. Light microscopy identified several coelomocyte types, including petaloid, filopodial, vibratile cells, and spherulocytes. The HCCM medium supplemented with coelomic fluid proved most effective for cell growth and viability. Moreover, coelomic fluid is the best culture media for differentiation of coelomocyte into the cell producing naphthoquinone pigments. These findings contribute to developing in vitro cell culture methods for sea urchin, providing a foundation for further research on sea urchin immunology, cell biology, and cellular responses to pathogens and other biological stress.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"863-874"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145389079","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 insect research through cell line transcriptomics.","authors":"Surjeet Kumar Arya, Subba Reddy Palli, Cynthia L Goodman","doi":"10.1007/s11626-026-01155-1","DOIUrl":"10.1007/s11626-026-01155-1","url":null,"abstract":"<p><p>This review emphasizes the significance of insect cell lines in transcriptomic research, highlighting their role as vital tools for uncovering cellular and molecular mechanisms of insect physiology, immune responses, and adaptation to environmental stressors. Cell lines derived from tissues such as the midgut, fat body, nervous system, and reproductive organs enable researchers to examine gene expression changes in a controlled setting, making discoveries that are difficult to achieve through whole-organism studies. High-throughput sequencing and single-cell RNA sequencing (scRNA-seq) have identified genes linked to detoxification, stress response, development, and immune defense, offering valuable insights for future applications in agriculture, pest control, and biotechnology. To organize this information clearly, we have summarized key findings in a table, providing an accessible overview of each cell line's important roles in transcriptomic research. This method not only highlights the adaptability of insect cell lines in functional genomics but also underscores their usefulness as model systems in pest management, virology, and bioengineering. Through utilizing transcriptomics, insect cell lines continue to advance our understanding of insect biology and foster the development of innovative strategies for sustainable crop protection and biotechnological use.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"937-951"},"PeriodicalIF":2.0,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146179508","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":"Role of GPCR41 silencing in conjugated linoleic acid-induced adipogenesis of bovine intramuscular preadipocytes.","authors":"Rong Wan, Chuanfang Zhang, Qingxiang Meng, Siwei Nong, Zhenming Zhou","doi":"10.1007/s11626-026-01209-4","DOIUrl":"https://doi.org/10.1007/s11626-026-01209-4","url":null,"abstract":"<p><p>The present study examined the impact of conjugated linoleic acid (CLA) on the differentiation and proliferation of bovine intramuscular preadipocyte (BIP) cells, specifically exploring the role of G protein-coupled receptor 41 (GPCR41). Cells were exposed to CLA at various concentrations ranging from 50 to 150 μmol/L. Flow cytometry revealed that treatment with 100 μmol/L CLA led to a significant increase in the proliferation rate. During differentiation, 100 and 150 μmol/L CLA markedly enhanced lipid droplet accumulation. Maximal cytoplasmic lipid content was observed when CLA was combined with insulin and dexamethasone. CLA treatment significantly enhanced the transcript abundance of the adipogenic transcription factors peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT-enhancer binding protein alpha (C/EBPα). Knockdown of GPCR41 via RNA interference in CLA-treated cells led to a significant suppression of GPCR41 expression, accompanied by decreased PPARγ mRNA levels and diminished cytoplasmic lipid droplet formation, indicating impaired differentiation. Taken together, these results show that CLA enhances both differentiation and proliferation of bovine intramuscular preadipocytes, possibly via activation of C/EBPα and PPARγ, with GPCR41 playing a critical role in mediating this adipogenic effect.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":2.0,"publicationDate":"2026-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148648361","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":"Endothelial cell-derived microRNAs-containing extracellular vesicles and diabetic retinopathy in a mouse model of diabetes.","authors":"Meili Wu, Haoyuan Zhou, Tianyi Zong, Ziwen Li, Yujuan Cao, Qian Yang, Tong Mu, Tianhua Xie, Xiaolu Wang, Yong Yao","doi":"10.1007/s11626-026-01205-8","DOIUrl":"https://doi.org/10.1007/s11626-026-01205-8","url":null,"abstract":"<p><p>Diabetic retinopathy (DR) is a leading cause of visual impairment and blindness in industrialized countries, resulting from diabetes mellitus. Prostaglandin E₂ (PGE₂), synthesized by cyclooxygenases, contributes to inflammation and apoptosis via the E-prostanoid receptor 2 (EP₂R). Our previous studies demonstrated that EP₂R antagonists mitigate inflammation and microvascular dysfunction in streptozotocin (STZ)-induced DR. Given the paracrine role of extracellular vesicles (EVs) in DR, we hypothesized that EVs derived from human endothelial cells (ECs) may regulate the PGE₂/EP₂R pathway in DR. Using an STZ-induced diabetic mouse model, we administered intravitreal injections of AAV2-shEP₂R and evaluated retinal histology, optical coherence tomography, and biochemical markers. EV morphology, size, and concentration from high glucose (HG)-treated ECs were analyzed. Small RNA expression in plasma EVs from DR patients was assessed via deep sequencing. EP₂R inhibition via AAV2-mediated knockdown significantly reduced retinal vascular leakage, leukostasis, and retinal Müller cell (rMC) activation. MiRNA profiling revealed elevated levels of miR-423-5p and miR-21-5p in EVs from HG-treated ECs, which were suppressed in EVs from EP₂R antagonist-treated cells. Notably, deep sequencing of plasma EVs from DR patients confirmed significant upregulation of these miRNAs compared to healthy controls. MiR-423-5p and miR-21-5p function as key paracrine mediators promoting Müller cell activation and retinal microvascular dysfunction in DR. These findings highlight the potential of circulating EVs as vehicles for miRNA-based therapeutic interventions in DR.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":2.0,"publicationDate":"2026-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148619996","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}