In Vitro Cellular & Developmental Biology. Animal最新文献

{"title":"A comparative genotoxicity study of agrochemicals: nuclear abnormalities, comet assay, and gene expression alterations.","authors":"Ankita Salunke, Parth Pandya, Bhumi Thakkar, Pragna Parikh","doi":"10.1007/s11626-025-01030-5","DOIUrl":"https://doi.org/10.1007/s11626-025-01030-5","url":null,"abstract":"<p><p>Agrochemicals (AGs) are known for their ability to have a negative impact on the health of non-target species, despite the fact that they are meant to protect agricultural plants from harmful pests. Catla catla (Hamilton, 1822) gill cells (ICG) were exposed to four AGs: insecticide (Imidacloprid (IMI)), fungicide (Curzate (CZ)), herbicide (pyrazosulfuron ethyl (PE)), and fertilizer micronutrients (MN) with sublethal concentrations 1/20th, 1/10th, and 1/5th of IC₅₀, described here as low dose (LD), medium dose (MD), and high dose (HD), respectively. A significant dose-dependent increase in the nuclear abnormalities such as micronuclei formation, bi-nucleated, and lobbed nucleated cells was observed in ICG cells treated with AGs. Of all the AGs, maximum alterations were observed with the HD of IMI followed by CZ, PE, and MN. Concurrently, the genotoxicity was determined by performing comet assays with high dose of all AGs. The gene expression of dnmt and cyp p450 were also studied through q-PCR in ICG cells. The significant increase in expression as well as alteration in cyp p450 and dnmt sequence was reported in ICG cells exposed to HD of IMI. This suggests that IMI has a genotoxic effect and may lead to epigenetic alterations.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008886","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":"Establishment of an embryonic cell line of Grapholita molesta (Lepidoptera: Tortricidae) and in vitro replication of Cydia pomonella granulovirus in it.","authors":"Gui-Ling Zheng, Yu-Fan Yao, Xiao-Yu Zhang, Qian-Long Yu, Jie Li, Yi-Ping Li, Dong Chu, Chang-You Li","doi":"10.1007/s11626-025-01036-z","DOIUrl":"https://doi.org/10.1007/s11626-025-01036-z","url":null,"abstract":"<p><p>The oriental fruit moth, Grapholita molesta (Busck) (Lepidoptera: Tortricidae), is a major pest of fruit trees worldwide. In this study, an embryonic cell line QAU-Gm-E-L of the oriental fruit moth was successfully established. The cells grew adherently, round cells and spindle cells accounted for 43.0% and 42.2% of the total population, respectively, and rod-shaped cells accounted for 14.8%. The amplified mitochondrial cytochrome oxidase I subunit (CoI) gene fragment was 651 bp in length, and its similarity with the CoI gene of the oriental fruit moth was 100%. The chromosomes of QAU-Gm-E-L cells were granular or short rod-shaped. Its number varied from 66 to 444, indicating that aneuploidy occurred. The observations were consistent with the chromosome characteristics of lepidopteran insect cell lines. The population doubling time of QAU-Gm-E-L cells was 27.64 h. Real-time fluorescence quantitative polymerase chain reaction (qPCR) confirmed that the number of copies of Cydia pomonella granulovirus (CpGV) gradually increased in QAU-Gm-E-L cells with inoculation time. The electron microscopy observations results showed that occlusion bodies (OBs) of CpGV could be formed in the cells at 4 d post-infection; a large number of OBs were seen in the cells at 8 d post-infection. Hence, the QAU-Gm-E-L cells can support the in vitro replication and proliferation of CpGV, and it will provide an ideal material for the molecular biology research of oriental fruit moth and CpGV.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012807","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":"Improve your success with fish cell lines-small things that matter.","authors":"Anita Solhaug, Georgina C Dowd, Vivian R Dayeh, Hilde Sindre, Lucy E J Lee, Niels C Bols","doi":"10.1007/s11626-025-01042-1","DOIUrl":"https://doi.org/10.1007/s11626-025-01042-1","url":null,"abstract":"<p><p>There is a drive towards reducing animal experiments and developing robust biologically relevant in vitro models based on cell lines, including those derived from fish. At the time of writing, Cellosaurus, the knowledge base of current cell lines used in research, listed more than 900 fish cell lines in its database. One of the key challenges facing fish cell biology is the lack of fundamental technical information regarding the isolation, culture, and application of cell lines. Researchers often work in silos, encountering similar technical challenges, each spending significant time and resources overcoming the same issues for which solutions may not be readily accessible. Here, we share some of the key considerations for the isolation, culture, maintenance, and application of fish cell lines in toxicology, which we have encountered over our collective decades of experience.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of small-scale laboratory-grown cell-based fish meat from Asian seabass muscle and fin cell lines.","authors":"Sivaraj Mithra, Seepoo Abdul Majeed, Shaik Abdullah Eisa Abdullah, Ganesan Ajay Pathra, Gani Taju, Isaac Sarojini Bright Singh, Perumal Santhanam, Azeez Sait Sahul Hameed","doi":"10.1007/s11626-025-01040-3","DOIUrl":"https://doi.org/10.1007/s11626-025-01040-3","url":null,"abstract":"<p><p>Aquaculture is essential to satisfying the world's increasing demand for seafood. Likewise, overfishing is becoming more common across the world, inflicting tremendous damage to the marine environment. There is a critical need for protecting sustainable fishing resources to fulfil the increasing demand for seafood. The current work focuses on the cells derived from Asian seabass muscle (SBM) and Asian seabass fin (SBF) for producing cell-based fish meat. SBM and SBF cells were seeded separately in the TubeSpin bioreactor and placed on a 3D orbital rocker. Cell sheets formed on the TubeSpin were detached and formed spheroid-like structures. These structures aggregated and formed visible tissue-like structures on 45 d of culture. Immunotyping results revealed that the presence of myosin in the cells of muscle and fin tissue, and indicating that these cells might have originated from myoblasts. The origin of cultured tissue from SBM and SBF cell lines was confirmed by amplification and sequencing of the L. calcarifer specific mitochondrial larger subunit rRNA gene. Additionally, these cells could be cultivated in multilayered forms that were appropriate for large-scale production. This approach provides a new method for the production of cell-based, laboratory-grown meat from the Asian seabass muscle and fin cell lines.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143811265","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":"Establishment of 27 cell lines derived from various insects.","authors":"Kazuyo Watanabe, Shigeo Imanishi, Takumi Kayukawa, Ken Tateishi","doi":"10.1007/s11626-025-01031-4","DOIUrl":"https://doi.org/10.1007/s11626-025-01031-4","url":null,"abstract":"<p><p>Insect cell lines are valuable for basic and applied biological research. In this study, we established 27 cell lines from various insect species, including Hemiptera: Nilaparvata lugens, Coleoptera: Sitophilus oryzae, Hymenoptera: Allantus luctifer and Trichogramma ssp., Diptera: Culicoides oxystoma, Lepidoptera: Spodoptera litura, Mythimna separata, Bombyx mori, Agrius convolvuli, Plodia interpunctella, and Cryptophlebia horii. This is the first report of cell lines derived from A. luctifer, C. oxystoma, A. convolvuli, and C. horii. Additionally, cell lines from S. litura and M. separata were established from different tissues including the hemocytes, fat bodies, embryos, and Malpighian tubules. Eighteen cell lines were successfully adapted to commercial culture media, with the population doubling time ranging from 1 to 8 d. The identities of the cell lines were confirmed using DNA barcoding. These established cell lines could be valuable for various research applications.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":""},"PeriodicalIF":1.5,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143811264","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":"Epidermal growth factor increases cystathionine β-synthase expression in cultured embryonic spinal cord cells.","authors":"Ryota Eguchi, Yuya Higashida, Mizuki Oouchi, Soichiro Yamaguchi, Ken-Ichi Otsuguro","doi":"10.1007/s11626-025-01043-0","DOIUrl":"10.1007/s11626-025-01043-0","url":null,"abstract":"<p><p>In the central nervous system (CNS), cystathionine β-synthase (CBS) is localized in astrocytes. CBS degrades cytotoxic homocysteine and produces cytoprotective hydrogen sulfide; thus the proper expression of CBS is required to maintain CNS functions. CBS expression is very low at the late embryonic stage and increases after birth. This study examined CBS expression in cultured spinal cord cells derived from fetal rats. Treatment of spinal cord cells with epidermal growth factor (EGF) promoted the proliferation and maturation of astrocytes during development. EGF (30 ng/ml, 4 days) increased CBS protein expression and the number of CBS-expressing astrocytes in the culture. A high cell density also increased CBS expression, and EGF was able to increase CBS expression when cellular proliferation was inhibited. The EGF receptor was predominately expressed in neural stem cells rather than astrocytes. These results suggest that EGF acts on neural stem cells, leading to increase in CBS-expressing astrocytes. This effect may reflect the maturation process of astrocytes during embryonic development.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"416-424"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144077790","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":"Neuroprotective effects of human umbilical cord mesenchymal stem cells (Neuroncell-EX) in a rat model of ischemic stroke are mediated by immunomodulation, blood-brain barrier integrity, angiogenesis, and neurogenesis.","authors":"Sze-Piaw Chin, Erlena Nor Asmira Abd Rahim, Natasha Najwa Nor Arfuzir","doi":"10.1007/s11626-025-01037-y","DOIUrl":"10.1007/s11626-025-01037-y","url":null,"abstract":"<p><p>Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are a potential off-the-shelf product for acute ischemic stroke. This study explored the underlying mechanism of Cytopeutics® hUC-MSCs (Neuroncell-EX) as well as its feasibility and efficacy at two different doses: 2 × 10⁶ cells per rat and 4 × 10⁶ cells/rat in middle cerebral artery occlusion (MCAO) ischemic stroke model for 28 d. Modified neurological severity score (mNSS) and rotarod tests were evaluated at days 1, 4, 7, and 14. Transforming growth factor-beta 1 (TGF-β1), interleukin-1 receptor antagonist (IL-1Ra), and vascular endothelial growth factor (VEGF) were evaluated by enzyme-linked immunosorbent assay (ELISA) at days 4 and 28. Immunohistochemistry expression of aquaporin-4 (AQP4) and neuronal protein marker (NeuN) were performed at days 4 and 28, respectively. Both doses of Neuroncell-EX showed significant lower mNSS scores at days 7 and 14 compared to stroke control. Both Neuroncell-EX groups showed significant longer latency time at day 7, with only 4 × 10⁶ cells/rat group having significant longer time at day 14 than stroke control. At both time points, the 2 × 10⁶ cells/rat group had significantly higher TGF-β1 and IL-1Ra levels, with significantly increased TGF-β1 only observed in 4 × 10⁶ cells/rat group at day 4 compared to stroke control. The VEGF levels were significantly lower at day 4 but then significantly increased at day 28 in both Neuroncell-EX groups than stroke control. AQP4 expression was significantly higher in stroke control compared to healthy control at day 4. Both doses of Neuroncell-EX showed significantly higher NeuN expression compared to stroke control at day 28. There is a weak correlation between TGF-β1 with VEGF and inversely with AQP4. These results suggest that Neuroncell-EX is feasible and effective in promoting functional recovery and neuroprotection in ischemic rats, potentially through immunomodulation, angiogenesis, and neurogenesis mechanisms.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"389-402"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12125091/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143963456","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 and characterization of canine umbilical cord mesenchymal/stromal stem cells.","authors":"Aline Pimentel, Triciana Gonçalves-Silva, Jasmin, Rosalia Mendez-Otero","doi":"10.1007/s11626-025-01023-4","DOIUrl":"10.1007/s11626-025-01023-4","url":null,"abstract":"<p><p>Mesenchymal stem cells (MSCs) have therapeutic potential due to their immunomodulatory and anti-inflammatory properties. In veterinary medicine, adipose tissue is the most common source of MSCs to treat canine disease, but the collection process is invasive, and the cells are influenced by the age and health conditions of the donor. These problems enhance interest in seeking alternative MSC sources, such as perinatal tissues. In this study, we developed and validated an optimized protocol for isolating canine umbilical cord MSCs for application in veterinary therapies. Umbilical cords obtained from cesarean sections were processed using three different protocols, involving combinations of mechanical and enzymatic tissue dissociation. The cells were cultured and evaluated for membrane receptors by flow cytometry to identify MSCs and assessed for their differentiation capacity. The number of cells obtained did not differ significantly between the combined protocol with trypsin and collagenase (TRIP + COL) and the collagenase protocol (COL). In in vitro culture, the combined TRIP + COL and COL yielded 12 to 14 times more cells, respectively, in the first passage than the explant (EXP) group, within fewer days of culture. Additionally, the cells obtained from these protocols showed a greater capacity for expansion over passages, and cells from both protocols showed fibroblast-like morphology and proliferation capacity up to the sixth passage. The cells obtained from these protocols were characterized by phenotype: CD45^-, CD34^-, CD14^-, HLA-DR^-, CD29⁺, CD44⁺, and CD90⁺, consistent with MSC identity. However, CD90 expression in the cells decreased significantly at sixth passage. Regarding differentiation, cells obtained from the COL protocol showed a capacity for commitment to the chondrogenic and osteogenic lineages. In conclusion, the COL and TRIP + COL protocols were more effective than the EXP protocol in terms of both the number and quality of isolated cells. However, due to its less-aggressive enzymatic nature, we considered the COL protocol to be the best method to obtain canine MSCs.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"472-485"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143998457","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":"(-)-Epicatechin regulates the resistance of lung adenocarcinoma cells to radiotherapy through the downregulation of FOXM1.","authors":"Jie Xia, Hongying Xu, Sihan Zhou, Tianqian Li, Zengbo Lv, Yingyu Yang, Meifang Huang","doi":"10.1007/s11626-025-01038-x","DOIUrl":"10.1007/s11626-025-01038-x","url":null,"abstract":"<p><p>Radioresistance, particularly as manifested by cancer stem cells (CSCs), is the most common reason for the failure of cancer radiotherapy. It is essential for effective radiotherapy to inhibit cancer cell stemness. Research indicates that (-)-epicatechin (EC) enhances the radiosensitivity of non-small cell lung cancer (NSCLC); however, its influence on cell stemness in lung adenocarcinoma (LUAD) resistant to radiotherapy is still not well understood. In this study, radioresistant cell lines A549R and H1299R were constructed by repeatedly irradiating A549 and H1299 cells with gradient doses of X-rays. CCK-8, cell cloning, flow cytometry, RT-qPCR, Western blot, sphere formation detection, and other methods were used for experimental exploration. This study revealed that the radioresistance of LUAD cells was related to their stemness. By inhibiting KLF4, SOX2, CD133, and ALDH1A1 expression, EC treatment increased radiosensitivity and reduced cell sphere formation. Also, FOXM1 expression was upregulated in LUAD and in radioresistant LUAD cells. Knocking down FOXM1 inhibited the stemness of radioresistant LUAD cells. Mechanistically, EC inhibited radiotherapy-resistant LUAD cell stemness by downregulating FOXM1 expression, thereby increasing radiosensitivity. In summary, our study revealed that EC inhibited radiotherapy resistance in LUAD cells through downregulating FOXM1, and it provides a theoretical framework for treating LUAD clinically.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"438-449"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144005282","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":"Polyvinyl alcohol can replace the fetal bovine serum during cryopreservation of canine adipose mesenchymal stromal cells.","authors":"Gabriel Corrêa de Camargo, Fernanda da Cruz Landim-Alvarenga, Alice Pereira Maciel, Driéle Bretones Dos Santos, Camilla de Paula Freitas Dell'Aqua, Marina Landim E Alvarenga, Amanda de Barros Piffer, Fabiana Ferreira de Souza","doi":"10.1007/s11626-025-01046-x","DOIUrl":"10.1007/s11626-025-01046-x","url":null,"abstract":"<p><p>Mesenchymal stromal cells (MSCs) are cells with multipotent characteristics present in various tissues and used as a promising alternative in cell therapy protocols in animals and humans. Creating stem cell banks for various purposes through cryopreservation is a common practice with MSCs. In this regard, the association between 10% dimethyl sulfoxide (Me2SO) and 90% fetal bovine serum (FBS) is widely used as a cryoprotective protocol for MSCs. However, these components have disadvantages, with possible risks to therapy receivers, contamination, and cytotoxic effects on MSCs. To replacing and reducing the use of FBS in the MSCs cryopreservation protocols, four agents were selected, being FBS at 10%, methylcellulose (MC) at 0.1%, polyvinyl alcohol (PVA) at 1%, and bovine albumin (BSA) at 1%, all associated with 10% Me2SO and 80% DMEM high glucose media. In the cell viability test with flow cytometry, the group with MC at 0.1% performed significantly worse than other treatments, except for BSA, which had a similar performance to MC. The expression of membrane proteins evaluation with flow cytometry showed that the cells treated with PVA and 10% FBS performed better at expressing lower values of CD34 and MHC-II. There were no differences regarding the osteogenic and adipogenic differentiation induction between the groups. We concluded that low concentrations of FBS (10% in DMEM) associated with BSA or PVA have a similar protective effect on cell viability during cryopreservation with Me2SO as 90% FBS. PVA showed an additional effect since the MSCs expressed lower concentrations of MHC-II and CD34.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"369-373"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144119565","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}