Cytotechnology最新文献

{"title":"Functional changes in β-lactoglobulin by conjugation with carboxymethyl cellulose.","authors":"Tatsuya Arai, Moeko Ono, Maiko Yoneda, Marika Sugamura, Tadashi Yoshida, Makoto Hattori","doi":"10.1007/s10616-025-00741-7","DOIUrl":"10.1007/s10616-025-00741-7","url":null,"abstract":"<p><p>β-lactoglobulin (BLG) and carboxymethylcellulose (CMC) were conjugated by using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC). The BLG-CMC conjugates with different CMC content and molecular weights were prepared. Confirmation of conjugation was carried out by SDS-PAGE. CD spectra revealed that the secondary structure of BLG had maintained in the conjugates. Fluorescence studies indicated that the conformation around Trp had not changed in the conjugates. Retinol-binding activity indicated that the retinol-binding site of BLG changed by the conjugation. Equilibrium constants (K_AS) of anti-BLG monoclonal antibodies (mAbs) to BLG after conjugating with CMC by competitive ELISA indicated that the structure around ¹⁵Val-²⁹Ile and ⁸Lys-¹⁹Trp maintained their native structure, and the structure around ¹²⁵Thr-¹³⁵Lys changed by conjugation. By conjugation with CMC, emulsifying property of BLG in the acidic pH region and in the presence of NaCl were much improved. Because acidic pH and salt are frequently used in food, the BLG-CMC conjugates are considered to be useful for food applications. Immunogenicity of BLG in BALB/c mice was reduced by this conjugation. In particular, there was a marked improvement in both emulsifying property and reduced immunogenicity in the BLG-high molecular weight (HMW) CMC conjugate. Therefore, conjugation with CMC is an effective way to improve BLG's function, and CMC with a high molecular weight is preferable.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 3","pages":"79"},"PeriodicalIF":2.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11910463/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143647329","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":"Blocking circular RNA FNDC3B induces fibroblast-like synoviocytes dysfunction to ameliorate rheumatoid arthritis through regulating the miR-125a-5p-Hexokinase2 axis.","authors":"Jiaxin Fu, Zhi Liu, Guangxin Zhang, Chun Zhang","doi":"10.1007/s10616-025-00745-3","DOIUrl":"10.1007/s10616-025-00745-3","url":null,"abstract":"<p><p>Rheumatoid arthritis (RA) is a chronic, progressive, autoimmune inflammatory joint disease. The cause of synovitis in rheumatoid arthritis involves the interaction between immune cells/macrophages and fibroblast-like synoviocytes (FLSs-RA). The impact of circular RNAs on FLSs and their role in RA pathology is still unknown. This study aimed to investigate the roles and molecular mechanisms of circular RNA FNDC3B in regulating cell injury and glucose metabolism of FLSs in RA. We demonstrated that circFNDC3B was significantly upregulated and miR-125a-5p was significantly downregulated in FLSs from RA patients. When circFNDC3B was silenced or miR-125a-5p was overexpressed, it reduced FLSs-RA glucose metabolism and increased oxidative stress-induced cell injury. Through bioinformatics analysis, RNA pull-down, and luciferase assays, it was found that circFNDC3B sponged miR-125a-5p to create a ceRNA network in FLSs-RA. The glucose metabolism rate was elevated in FLSs-RA, showing a glucose-dependent characteristic compared to normal FLSs. The enzyme hexokinase 2 (HK2), which is crucial for glucose metabolism, was identified as a direct target of miR-125a-5p in FLSs. In rescue experiments, restoring miR-125a-5p in circFNDC3B-overexpressing FLSs-RA successfully counteracted the circFNDC3B-promoted glucose metabolism and resistance to cell injury. In conclusion, this study highlighted the important roles and molecular mechanisms of circFNDC3B in accelerating glucose metabolism and preventing cell apoptosis in fibroblast-like synoviocytes during rheumatoid arthritis by modulating the miR-125a-5p-HK2 axis. Targeting the circFNDC3B-mediated glucose metabolism pathway could be a promising strategy for rheumatoid arthritis therapy.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00745-3.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 3","pages":"83"},"PeriodicalIF":2.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11947371/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143751456","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":"Exploring the mechanism of <i>Epimedium</i> in treating diabetic nephropathy based on network pharmacology and experimental validation study.","authors":"Leyu Huang, Hui Li, Ying Han","doi":"10.1007/s10616-025-00748-0","DOIUrl":"10.1007/s10616-025-00748-0","url":null,"abstract":"<p><p>Diabetic nephropathy (DN) is a severe complication of diabetes, characterized by chronic inflammation, metabolic disturbances, and progressive renal damage. Natural perennial herb, such as <i>Epimedium</i>, has shown potential therapeutic effects on DN, but its underlying mechanisms remain unclear. This study aimed to explore the pharmacological mechanisms of <i>Epimedium</i> in the treatment of DN through network pharmacology, molecular docking, and experimental validation. Active components of <i>Epimedium</i> were identified using TCMSP and SwissTargetPrediction databases, while DN-related targets were retrieved from GeneCards, DisGeNET, OMIM, and TTD databases. Overlapping targets were analyzed via PPI network and Cytoscape's cytoHubba plugin to identify hub genes. GO and KEGG enrichment analyses were conducted to explore functional pathways. Molecular docking validated the binding affinity between key targets and active components. Finally, high-glucose-induced HK-2 cell injury models were used to verify the protective effects of <i>Epimedium</i> through RT-qPCR, western blotting, and mitochondrial function assays. A total of 224 overlapping targets were identified, with <i>AKT1, TNF, HSP90AA1</i>, and <i>SRC</i> serving as key hub genes. GO and KEGG analyses revealed significant enrichment in pathways such as the PI3K-Akt signaling pathway and lipid metabolism. Molecular docking demonstrated strong interactions between <i>Epimedium</i> components and hub targets. Experimental validation showed that <i>Epimedium</i> restored nephrin and WT1 protein levels, mitigated mitochondrial dysfunction, and reversed high-glucose-induced overexpression of key targets. <i>Epimedium</i> exerts therapeutic effects on DN through multi-target interactions, primarily via the PI3K-Akt pathway, highlighting its potential as a novel treatment for DN.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00748-0.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 3","pages":"82"},"PeriodicalIF":2.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11937453/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143729207","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":"Astragalin inhibits the proliferation of high-risk HPV-positive cervical epithelial cells and attenuates malignant cervical lesions.","authors":"Wei Zeng, Li Chen","doi":"10.1007/s10616-025-00742-6","DOIUrl":"10.1007/s10616-025-00742-6","url":null,"abstract":"<p><p>High-risk human papillomavirus (HPV), especially HPV16 and HPV18, are closely linked to the onset of cervical cancer (CC). Astragalin (AST), a bioactive flavonoid, has been reported to impede CC HeLa cell proliferation. Nevertheless, the mechanism by which AST exerts its tumor-suppressive role in CC remains unclear. HeLa (HPV18-positive) and CaSki (HPV16-positive) cells were exposed to various concentrations of AST. CCK-8 assay, flow cytometry analysis, wound healing, and Transwell assays were employed to examine the AST functions on CC cell aggressiveness. Protein levels were assessed by western blotting. Immunofluorescence staining was used to detect E6, E7, p53, and p-pRb expression. Animal experiments were performed to validate the anti-CC role in vivo. The results showed that AST dose-dependently impaired HeLa and CaSki cell viability and elicited G1 cell cycle arrest. AST restrained CC cell migration and invasiveness. AST inhibited the growth of HeLa-derived xenograft tumors in mice and repressed E6/E7 oncoprotein expression in CC cells and mouse tumor tissues. In conclusion, AST suppresses CC progression by downregulating E6/E7 oncoprotein expression to attenuate CC cell aggressiveness.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00742-6.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 3","pages":"80"},"PeriodicalIF":2.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11923324/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691479","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":"Effect of different cell culture media on the production and glycosylation of a monoclonal antibody from a CHO cell line.","authors":"Jaeweon Lee, Uriel Ortega-Rodriguez, Chikkathur N Madhavarao, Tongzhong Ju, Thomas O'Connor, Muhammad Ashraf, Seongkyu Yoon","doi":"10.1007/s10616-025-00733-7","DOIUrl":"10.1007/s10616-025-00733-7","url":null,"abstract":"<p><p>Recombinant monoclonal antibodies (mAbs) are commonly produced using Chinese hamster ovary (CHO) cells and the cell culture medium used in bioreactors influences the yield and quality attributes of the protein drug products. The COVID 19 pandemic revealed a vulnerability in the supply chain for necessary reagents (such as culture medium and raw material) for maintaining un-interrupted production of protein drugs with consistent quality. The supply interruption for the cell culture medium ActiPro™ optimized for producing VRC01, an IgG1-κ mAb, from a CHO-K1 cell line, necessitated the search for alternate media. VRC01 mAb is highly glycosylated and can broadly neutralize several strains of Human Immunodeficiency Virus (HIV). We investigated to see if an alternate medium can be used in the production without impacting quality attributes like glycosylation. In our strategy, we used 3 different commercially available media, performed two sets of experiments-with and without media supplements, Cell boost 7a and Cell boost 7b. Cell growth, volumetric production of the mAb protein and glycosylation pattern were compared to identify an alternative medium. Among the tested media based on cell growth, mAb production potential and glycosylation analysis, ActiCHO™ P was found to be a better alternate medium to ActiPro™ medium than EX-CELL® 325 PF CHO medium to produce VRC01 mAb. Overall, the approach used here to establish the impact of variation in medium on protein therapeutic attributes may be used during product development to build in supply chain resilience in drug manufacturing.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00733-7.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 3","pages":"81"},"PeriodicalIF":2.0,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11928345/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143691483","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":"Expression and role of CTHRC1 in inflammatory bowel disease in children.","authors":"Heng Tang, Xiang Gao, Zhaofang Wu, Jia Chen, Li Chen, Xiang Du","doi":"10.1007/s10616-025-00705-x","DOIUrl":"10.1007/s10616-025-00705-x","url":null,"abstract":"<p><p>Inflammatory bowel disease (IBD) is a chronic, progressive, immune-mediated, gastrointestinal inflammatory disease with increasing occurrences in children. Collagen triple helix repeat containing 1 (CTHRC1), a migration-promoting protein, acts as a tumor-promoting factor in malignant tumors. However, functions and mechanisms of CTHRC1 in children with IBD remain unclear. This study aimed to determine the effects and mechanisms of CTHRC1 on dextran sodium sulfate (DSS)-treated HT-29 cells. HT-29 control cells were exposed to 2% DSS to develop an in vitro IBD model. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to assess CTHRC1 expression in serum of children with IBD and HT-29 cells. Cell viability and apoptosis were assessed using MTT and flow cytometry (FCM). Expressions of cleaved-Caspase3 and Caspase3 were determined by western blotting. The cytokine production (TNF-α, IL-1β and IL-6) in HT-29 cells was measured by ELISA assay. Activation or inactivation of NF-κB signaling pathway was confirmed by western blot assay. Results showed that CTHRC1 expression was upregulated in the IBD serum and HT-29 control cells. The level of CTHRC1 was lower in CTHRC1-siRNA transfected cells than in control siRNA-treated cells. Notably, silence of CTHRC1 markedly enhanced HT-29 cells viability, decreased apoptotic cells, suppressed cleaved-Caspase3 expression, inhibited cleaved-Caspase3/Caspase3 ratio, reduced the production of inflammatory cytokines, and blocked NF-κB signaling pathway induced by DSS. However, these effects were reversed following diprovocim treatment. Thus, that knockdown of CTHRC1 alleviated DSS-induced HT-29 cell injury by inhibiting the NF-κB signaling pathway in vitro, providing a new therapeutic target for IBD in children.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00705-x.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 2","pages":"44"},"PeriodicalIF":2.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11759733/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143045406","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":"Incorporating shaken 24-deep-well plate fed-batch culture shortens CHO cell line development time.","authors":"Shunsuke Ohira, Takeshi Omasa","doi":"10.1007/s10616-025-00728-4","DOIUrl":"10.1007/s10616-025-00728-4","url":null,"abstract":"<p><p>In conventional Chinese hamster ovary (CHO) cell line development, static culture is used for early-stage screening, whereas suspension culture is generally used for the manufacturing process. Shaken flask (SF) fed-batch culture allows evaluation with culture mode, which is closer to the final process. However, due to its laborious and low-throughput characteristics, only a limited number of clones can be evaluated. To attain high-throughput fed-batch culture evaluation, a shaken 24-deep-well plate (24 DWP) culture process was developed. 24 DWP culture allows multiple plates to be run in parallel, and is therefore suitable for early-stage screening. One challenge of well plate culture is the nonuniform evaporation rate among wells, which may result in unnecessary bias on clone evaluation. The 'sandwich lid system' introduced here provides a uniform evaporation rate, and showed no significant difference in cell culture performance by well location. 192 antibody-producing CHO clones were evaluated by 24 DWP fed-batch culture, and 30 clones were selected. On comparison of clone sets selected by 24 DWP fed-batch culture and the conventional scheme, average antibody concentration in SF fed-batch culture was 3.6 g/L and 2.9 g/L, respectively. 24 DWP fed-batch culture process showed a high correlation ratio with SF fed-batch culture in antibody productivity and similar cell culture characteristics. These characteristics-high-throughput and sufficient culture volume to support cell culture performance monitoring-indicate that 24 DWP fed-batch culture can be applied in the clone selection stage in place of SF, and will shorten the time required for cell line development.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 2","pages":"64"},"PeriodicalIF":2.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11839547/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482474","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":"The roles of melatonin and potassium channels in relaxation response to ang 1-7 in diabetic rat isolated aorta.","authors":"Nazar M Shareef Mahmood, Almas M R Mahmood, Ismail M Maulood","doi":"10.1007/s10616-025-00720-y","DOIUrl":"10.1007/s10616-025-00720-y","url":null,"abstract":"<p><p>In a circadian cycle, the pineal gland produces and releases melatonin (MEL) into the bloodstream. By activating distinct melatonin receptors, MEL has been shown to variably change vascular endothelial dysfunction (VED) to various vascular beds. This study investigates the interaction of melatonin (MEL) and potassium ion (K⁺) on angiotensin 1-7 (Ang 1-7) vasorelaxant in streptozotocin (STZ)-induced diabetes mellitus (DM) and non-diabetes mellitus (non-DM) male albino rat aortic rings. The isometric tension of isolated aortic rings was assessed by generating a dose-response curve (DRC) for Ang 1-7 using a PowerLab data acquisition system. Accordingly, three experimental sets were carried out. In the first set the aortic rings were exposed MEL and MEL agonist ramelteon (RAM) and MEL antagonist luzindole (LUZ). In the second set, the aortic rings were exposed to various non-selective calcium activated potassium channel (K_Ca) blockers, including tetraethylammonium (TEA), a small and large-conductance calcium-activated K⁺ [(SK_Ca) and (BK_Ca)] channels blocker charybdotoxin (ChTx) and intermediate calcium-activated K⁺ channel (IK_Ca) blocker clotrimazole (CLT). In the third set, the aortic rings were exposed to various selective K⁺ channels blockers, including the selective blocker of K_ATP channel, glibenclamide (Glib), 4-aminopyridine (4-AP), a selective blocker of K_v channels and BaCl₂, delayed inward rectifier K⁺ channels (K_ir) blocker. The results highlight the significant role of MEL in modulating vascular reactivity, particularly in the DM aorta. By enhancing the vasorelaxant effects of Ang 1-7 through mechanisms involving its receptors and antioxidant activities, MEL demonstrates its potential to counteract oxidative stress and VED associated with diabetes. These findings advance the understanding of vascular reactivity in diabetes and suggest MEL as a promising therapeutic agent for improving vascular health in diabetic conditions.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 2","pages":"55"},"PeriodicalIF":2.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11799518/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143381846","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":"Dietary Zinc activates the Nrf2 signaling pathway to inhibit pyroptosis and attenuate the lung inflammatory response in COPD.","authors":"Yanqiu Huang, Tao Liang, Junfei Liu, Hongyan Yu, Jingna Li, Li Han","doi":"10.1007/s10616-025-00725-7","DOIUrl":"10.1007/s10616-025-00725-7","url":null,"abstract":"<p><p>Pyroptosis and inflammation play crucial roles in the development of chronic obstructive pulmonary disease (COPD), and Zinc deficiency is commonly observed in COPD patients. In this study, we aimed to explore the impact of Zinc supplementation on pyroptosis and inflammation in a cigarette smoke (CS)-induced COPD mouse model, as well as the underlying mechanisms. The COPD mouse model was established through CS exposure, and mouse pulmonary epithelial cells (MLE-12) were exposed to cigarette smoke extract (CSE) to further validate the effects of Zinc supplementation. CS exposure resulted in significant alveolar wall damage, increased thickening of the alveolar walls, and elevated levels of interleukin-1β (IL-1β), IL-6, IL-18, and tumor necrosis factor-α (TNF-α) in the lung tissues of COPD mice. However, treatment with dexamethasone (a positive control) or Zinc supplementation alleviated these damages. Furthermore, the expressions of pyroptosis markers, including NLRP3, cleaved-Caspase-1, and GSDMD-N proteins, were upregulated in the lung tissues after CS exposure. Zinc supplementation, however, reversed these changes. Additionally, Zinc supplementation upregulated the protein expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), and quinone oxidoreductase-1 (NQO-1), and promoted the ubiquitination of Kelch-like ECH-associated protein 1 (Keap1) mediated by tripartite motif 25 (TRIM25) in the lung tissues of CS-induced mice. Importantly, the Nrf2 signaling inhibitor ML385 abolished the beneficial effects of Zinc in CS-exposed mice. Similar results were observed in MLE-12 lung epithelial cells exposed to CSE. In summary, Zinc supplementation inhibits pyroptosis and attenuates inflammation in COPD mice by activating the Nrf2 pathway.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00725-7.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 2","pages":"62"},"PeriodicalIF":2.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11836256/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143467264","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":"RNA sequencing identifies <i>MAP1A</i> and <i>PTTG1</i> as predictive genes of aging CD264⁺ human mesenchymal stem cells at an early passage.","authors":"Margaret K Giler, H Alan Tucker, Amanda K Foote, Avery G Francis, Sean D Madsen, Yao-Zhong Liu, Kim C O'Connor","doi":"10.1007/s10616-025-00724-8","DOIUrl":"10.1007/s10616-025-00724-8","url":null,"abstract":"<p><p>Molecular profiles of mesenchymal stem cells (MSCs) are needed to standardize the composition and effectiveness of MSC therapeutics. This study employs RNA sequencing to identify genes to be used in concert with CD264 as a molecular profile of aging MSCs at a clinically relevant culture passage. CD264^- and CD264⁺ populations were isolated by fluorescence-activated cell sorting from passage 4 MSC cultures. CD264⁺ MSCs exhibited an aging phenotype relative to their CD264^- counterpart. Donor-matched CD264^-/+ mRNA samples from 5 donors were subjected to pair-ended, next-generation sequencing. An independent set of 5 donor MSCs was used to validate differential expression of select genes with quantitative reverse transcription PCR. Pairwise differential expression analysis identified 2,322 downregulated genes and 2,695 upregulated genes in CD264⁺ MSCs relative to donor-matched CD264^- MSCs with a Benjamini-Hochberg adjusted <i>p</i>-value (BH <i>p</i> _<i>adj</i> ) < 0.1. Nearly 25% of these genes were unique to CD264^-/+ MSCs and not differentially expressed at a significance level of BH <i>p</i> _<i>adj</i>  < 0.1 in previous RNA sequencing studies of early- vs. late-passage MSCs. Least Absolute Shrinkage and Selection Operator regression identified microtubule-associated protein 1A (<i>MAP1A</i>) and pituitary tumor-transforming gene 1 (<i>PTTG1</i>) as predictive genes of CD264⁺ MSCs. Combined <i>MAP1A</i> and <i>PTTG1</i> expression correctly classified CD264 status of MSC samples with an accuracy of 100%. Differential expression and predictive ability of <i>MAP1A</i> and <i>PTTG1</i> compared favorably with that of existing senescence markers expressed in early passage CD264^-/+ MSCs. This study provides the first linkage of <i>MAP1A</i> to CD264, aging and senescence. Our findings have application as quality metrics to standardize the composition of MSC therapies and as molecular targets to slow/reverse cellular aging.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s10616-025-00724-8.</p>","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"77 2","pages":"63"},"PeriodicalIF":2.0,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11839963/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143467289","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}