In Vitro Cellular & Developmental Biology. Animal最新文献

{"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}

{"title":"Correction: The adaptation of bovine embryonic stem cells to the changes of feeder layers.","authors":"Wenqiang Xu, Lingna Gao, Wei Li, Jing Wang, Yongli Yue, Xueling Li","doi":"10.1007/s11626-024-01013-y","DOIUrl":"10.1007/s11626-024-01013-y","url":null,"abstract":"","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"486-491"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143803117","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 a highly sensitive porcine alveolar macrophage cell line for African swine fever virus.","authors":"Xiangwan Lu, Xiadan Gong, Yingshuo Sun, Lang Gong, Yan Zhang","doi":"10.1007/s11626-025-01016-3","DOIUrl":"10.1007/s11626-025-01016-3","url":null,"abstract":"<p><p>African swine fever (ASF) caused by the African swine fever virus (ASFV) is a significant threat to domestic pig populations because of its highly contagious nature and associated morbidity and mortality. The lack of an appropriate cell line for ASFV propagation has significantly hindered the development of a safe and effective vaccine. In this study, we aimed to identify a cell line that is highly receptive to ASFV by evaluating various genes to determine their ability to support ASFV infection and replication. Our investigation revealed the efficient infection of a porcine alveolar macrophage cell line iPAM, upon stable overexpression of the transmembrane protein 107 (TMEM107). An isolated monoclonal cell line iPAM^{pCDH-TMEM107-B6} that was derived from the parental iPAM cell line exhibited increased susceptibility to ASFV infection. Notably, iPAM^{pCDH-TMEM107-B6} cells concurrently expressed ASFV B646L and ASFV p30 proteins after infection with ASFV. Biological characterization of iPAM^{pCDH-TMEM107-B6} revealed an enhanced proliferative capacity without compromised phagocytic function, indicating the retention of key cellular traits following genetic modification. The iPAM^{pCDH-TMEM107-B6} cell line has significant potential for ASFV research and will facilitate tasks such as isolation, replication, and genetic manipulation. The establishment of ASFV-sensitive cell lines provides an in vitro research platform for ASFV investigations, thereby advancing our understanding of the pathogenic mechanisms and aiding in vaccine development efforts.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"425-437"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143968654","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":"SPARC: a key mediator of apoptosis in human umbilical vein endothelial cells and its role in hypertension mechanism.","authors":"Yingyue Zhang, Haijing Zhao, Liuyang Tian, Zengao Yang, Li Zheng, Honghong Zhang, Yue Zhu, Yuhan Ma, Yong Xu, Yuqi Liu","doi":"10.1007/s11626-025-01026-1","DOIUrl":"10.1007/s11626-025-01026-1","url":null,"abstract":"<p><p>Hypertensionis a leading global health issue associated with high mortality and severe complications. Understanding its molecular mechanisms is essential for identifying novel therapeutic targets. Secreted protein acidic and rich in cysteine (SPARC) is associated with cell migration, disease pathophysiology, and inflammation; however, its role in hypertension remains under investigation. This study investigates the role of SPARC in hypertension, focusing on its impact on endothelial dysfunction.Using the GSE75815 dataset from the GEO database, we identified 71 differentially expressed genes (DEGs) associated with hypertension. Pathway analyses and protein-protein interaction networks constructed through the STRING database highlighted six hub genes, with further evaluation based on Comparative Toxicogenomics Database (CTD) scores. Immune cell profiling via ImmuCellAI revealed an increase in naive B cells, positively correlating with hub gene expression.Experimental validation in human umbilical vein endothelial cells (HUVECs) treated with angiotensin II demonstrated that SPARC downregulation reduced apoptosis and BAX expression. Silencing SPARC enhanced endothelial cell proliferation, migration, and nitric oxide production, counteracting angiotensin II-induced damage. Notably, angiotensin II upregulated SPARC secretion, suggesting its critical role in mediating endothelial dysfunction.These findings establish SPARC as a key contributor to the molecular pathways underlying hypertension. Targeting SPARC may represent a novel therapeutic strategy to mitigate endothelial dysfunction and improve outcomes for hypertensive patients.Our findings highlight SPARC as a key player in the molecular pathways of hypertension. Modulating SPARC expression may offer a promising therapeutic strategy to counteract endothelial dysfunction and improve outcomes in hypertensive patients.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"374-388"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143977852","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":"Overexpression of miR-20a targeting DUSP3 inhibits OCLN ubiquitination levels and alleviates sepsis induced intestinal barrier dysfunction.","authors":"Liming Cheng, Bo Feng, Chao Xie, Chunyan Chen, Linghui Guo","doi":"10.1007/s11626-025-01052-z","DOIUrl":"10.1007/s11626-025-01052-z","url":null,"abstract":"<p><p>Sepsis is a severe organ dysfunction syndrome caused by the host's dysfunctional response to infection. Sepsis can severely damage intestinal epithelial tissue, lead to intestinal barrier dysfunction, and seriously endanger human health. Therefore, this study aimed to explore the mechanism of miR-20a in sepsis-induced intestinal barrier dysfunction. In this study, mice and NCM460 cells were subjected to cecal ligation and puncture (CLP) and 1 μg/mL LPS, respectively, to establish a sepsis model. The expression of relevant genes, apoptosis, inflammation, and intestinal barrier dysfunction-related indices under the conditions of overexpression of miR-20a or DUSP3 and knockdown of DUSP3 or OCLN were assessed by western blotting, RT-qPCR, ELISA, flow cytometry, immunofluorescence, and HE staining. The experimental results revealed that in sepsis-induced intestinal barrier dysfunction, the expression of miR-20a and OCLN was downregulated, whereas that of DUSP3 was upregulated. Functionally, miR-20a inhibited LPS-induced intestinal epithelial cell apoptosis and inflammation and relieved sepsis-induced intestinal barrier dysfunction in mice. Experiments investigating the downstream mechanisms revealed that miR-20a overexpression suppressed LPS-induced intestinal epithelial cell apoptosis and inflammation and relieved sepsis-induced intestinal barrier dysfunction by targeting and inhibiting DUSP3 levels and OCLN ubiquitination. In conclusion, miR-20a relieves sepsis-induced intestinal barrier dysfunction by inhibiting DUSP3 and suppressing the ubiquitination of OCLN.</p>","PeriodicalId":13340,"journal":{"name":"In Vitro Cellular & Developmental Biology. Animal","volume":" ","pages":"459-471"},"PeriodicalIF":1.5,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144110575","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}