Cytotechnology最新文献_第9页

PRPF19 functions in DNA damage repair and gemcitabine sensitivity via regulating DDB1 in bladder cancer cells PRPF19通过调控膀胱癌细胞中的DDB1参与DNA损伤修复和吉西他滨敏感性

4区生物学

Cytotechnology Pub Date : 2023-10-05 DOI: 10.1007/s10616-023-00599-7

Jingjiang Yu, Shuxiong Ge

引用次数: 0

Assessing MTT and sulforhodamine B cell proliferation assays under multiple oxygen environments. 在多种氧气环境下评估MTT和磺基罗丹明B细胞增殖测定。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-07-05 DOI: 10.1007/s10616-023-00584-0

Ming Yao, Glenn Walker, Michael P Gamcsik

{"title":"Assessing MTT and sulforhodamine B cell proliferation assays under multiple oxygen environments.","authors":"Ming Yao, Glenn Walker, Michael P Gamcsik","doi":"10.1007/s10616-023-00584-0","DOIUrl":"10.1007/s10616-023-00584-0","url":null,"abstract":"Cell proliferation can be measured directly by counting cells or indirectly using assays that quantitate total protein or metabolic activity. However, for comparing cell proliferation under varying oxygen conditions it is not clear that these assays are appropriate surrogates for cell counting as cell metabolism and protein synthesis may vary under different oxygen environments. We used permeable bottom tissue culture ware to compare proliferation assays as a function of static oxygen concentrations under oxygen partial pressure (pO2) levels ranging from 2 to 139 mmHg. Cell proliferation was measured by cell counting and compared to surrogate methods measuring cell metabolism (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, MTT) and total protein (sulforhodamine B) assays under these different environments in Caco-2, MCF-7, MCF-10A and PANC-1 human cell lines. We found that the MTT readings do not correlate with cell number for the Caco-2 and PANC-1 cell lines under different oxygen conditions, whereas the sulforhodamine B protein assays perform well under all conditions. However, within a given oxygen environment, both proliferation assays show a correlation with cell number. Therefore, the MTT assay must be used with caution when comparing cell growth or drug response for cells grown in different oxygen environments.Supplementary information: The online version contains supplementary material available at 10.1007/s10616-023-00584-0.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"381-390"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465423/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10148801","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}

引用次数: 0

Effect assessment of a type of xeno-free and serum-free human adipose-derived mesenchymal stem cells culture medium by proliferation and differentiation capacities. 通过增殖和分化能力评估一种无异种和无血清的人脂肪来源的间充质干细胞培养基的效果。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-07-11 DOI: 10.1007/s10616-023-00586-y

Shanshan Chen, Li Meng, Shanshan Wang, Yan Xu, Wenbin Chen, Jianfeng Wei

{"title":"Effect assessment of a type of xeno-free and serum-free human adipose-derived mesenchymal stem cells culture medium by proliferation and differentiation capacities.","authors":"Shanshan Chen, Li Meng, Shanshan Wang, Yan Xu, Wenbin Chen, Jianfeng Wei","doi":"10.1007/s10616-023-00586-y","DOIUrl":"10.1007/s10616-023-00586-y","url":null,"abstract":"Human mesenchymal stem cells (hMSCs) possess broad prospects in pre-clinical research. In vitro amplification of hMSCs requires appropriate medium to reach the number of seed cells with clinical significance. However, the uncertainty of the heterologous components of the traditional fetal bovine serum (FBS) culture medium has great safety risks. Moreover, existing commercial hMSCs medium is very expensive, therefore a safer and more optimal hMSCs medium is urgently needed. Accordingly, we developed five components adipose-derived hMSCs (hADMSCs) medium without xenogenic components, named E5 SFM. which is mainly composed of knockout serum replacement (KSR), and additionally four components such as fibroblast growth factor and transferrin. Here, we mainly compared the E5 SFM with traditional FBS-containing medium and a commercial medium by surface markers testing, proliferation assay as well as osteogenic, adipogenic and chondrogenic differentiation assessment. We demonstrated that hADMSCs cultured in the E5 SFM showed similar morphological characteristics and immunophenotypes to those in other media. Notably, cell proliferative capability was similar to that in the commercial medium, but higher than that in the FBS-containing medium and other media. Additionally, their capabilities of adipogenic and osteogenic differentiation were significantly higher than those of other media. Consequently, we concluded that the E5 SFM medium can not only effectively promote cell proliferation of hMSCs, but also has optimal differentiative capacity and clear and simple ingredients.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"403-420"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465441/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137221","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}

引用次数: 0

H3K27 acetylation-induced FSTL1 upregulation by P300/RUNX1 co-activation exacerbated autophagy-mediated neuronal damage and NF-κB-stimulated inflammation in Alzheimer's disease. H3K27乙酰化诱导的FSTL1通过P300/RUNX1共激活上调，加剧了阿尔茨海默病中自噬介导的神经元损伤和NF-κB刺激的炎症。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-08-04 DOI: 10.1007/s10616-023-00589-9

Dongmei Dai, Junzheng Xie, Yun Zheng, Fangbin Chen, Bin Zhao, Li Miao

引用次数: 0

Effect of milk and whey on proliferation and differentiation of placental stromal cells. 牛奶和乳清对胎盘基质细胞增殖和分化的影响。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-07-17 DOI: 10.1007/s10616-023-00585-z

Bircan Boga, Merve Akbulut, Erkan Maytalman, Ilknur Kozanoglu

{"title":"Effect of milk and whey on proliferation and differentiation of placental stromal cells.","authors":"Bircan Boga, Merve Akbulut, Erkan Maytalman, Ilknur Kozanoglu","doi":"10.1007/s10616-023-00585-z","DOIUrl":"10.1007/s10616-023-00585-z","url":null,"abstract":"Fetal bovine serum (FBS), which is widely used in cell culture media, has the potential to cause medical and ethical problems. Here, an experimental study using milk or whey proteins containing essential nutrients and growth factors is presented to limit the use of FBS in cell culture media produced for cell and tissue regeneration. Study groups were formed by culturing human placenta mesenchymal stem cells, known to have high proliferation and differentiation capacity, with milk or whey solution at increasing concentrations, alone or in combination with FBS. Osteogenic and adipogenic differentiation capacities of proliferating cells were observed in FBS, milk or whey groups. Milk, whey or FBS groups obtained in P3 and after differentiation were separately analyzed for protein mRNA expression by reverse transcriptase-polymerase chain reaction (RT-qPCR). Fibroblast Growth Factor 2 (FGF2), Octamer-binding Transcription Factor 4 (OCT4), Bone Morphogenetic Protein 6 (BMP6), and adipogenic differentiation marker Peroxisome Proliferator-Activated Receptor Gamma (PPARG) were analysed by RT-qPCR. Proliferation was more pronounced in FBS alone and in its combinations with milk-whey compared to the groups in which only milk and whey were used. OCT4 mRNA and FGF2 mRNA expression decreased in differentiated cells. BMP6 mRNA expression increased with osteogenic and adipogenic stimuli. As expected, PPRG expression also increased with adipogenic stimulation. With this experimental study, evidence has been obtained that milk or whey can provide nutritional support to the culture media of repair cells and preserve the functional capacity of the cells, with a slightly more limited capacity than FBS.Graphical abstract: Supplementary information: The online version contains supplementary material available at 10.1007/s10616-023-00585-z.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"391-401"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465414/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137220","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}

引用次数: 0

Identification of differentially expressed miRNAs and mRNAs associated with the regulation of breast cancer via in silico and in vitro methods. 通过计算机和体外方法鉴定与癌症调节相关的差异表达miRNA和mRNA。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-07-03 DOI: 10.1007/s10616-023-00583-1

Pelin Telkoparan-Akillilar, Dilek Cevik

{"title":"Identification of differentially expressed miRNAs and mRNAs associated with the regulation of breast cancer via in silico and in vitro methods.","authors":"Pelin Telkoparan-Akillilar, Dilek Cevik","doi":"10.1007/s10616-023-00583-1","DOIUrl":"10.1007/s10616-023-00583-1","url":null,"abstract":"miRNA expressions are altered during development of breast cancer (BC). The aim of this study is to identify novel cancer-related miRNAs and pathways to understand the mechanisms of BC subtypes. GSE59247 dataset was downloaded from gene expression omnibus (GEO) database and analyzed with GEO2R software. The differential miRNA expressions in BC cells were evaluated by miRNome PCR array. Venn diagram was used to reveal co-differentially expressed miRNAs between GSE59247 dataset and miRNome array. Clinical prognostic significance of selected miRNAs was evaluated via Kaplan Meier curve. KEGG pathway enrichment analysis was performed to find miRNA targets and results were validated by TNM plot analysis and q-RT-PCR. TargetScan database was used to predict the association of miRNAs and 3'-untranslated regions of target genes and their expressions were visualized by human protein atlas database. Venn diagram analysis showed overlap of 11 miRNAs from in silico and in vitro analysis. KEGG analysis revealed 'Lysine Degradation Pathway' as the most significantly enriched targeted pathway. q-RT-PCR results confirmed that Lysine degradation pathway related genes SETD7, SETDB2, EHHADH, SETMAR, KMT2A and SUV39H2 were differentially expressed in BC cells. Target prediction analysis identified binding sites between miR-1323-5p and 3'-UTR of SETD7, miR-129-5p and 3'-UTR of EHHADH and miR-628-5p and 3'-UTR of SETDB2 mRNA. Notably, miR-1323-5p, miR-129-5p, and miR-628-5p are differentially expressed in BC and they bind to 3'UTR of critical genes of Lysine degradation pathway, namely SETD7, SETDB2 and EHHADH. These miRNAs might serve as potential diagnostic and prognostic biomarkers for progression.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"363-379"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465466/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10128100","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}

引用次数: 0

Lobetyolin inhibits cell proliferation and induces cell apoptosis by downregulating ASCT2 in gastric cancer. 洛贝托林通过下调癌症ASCT2抑制细胞增殖并诱导细胞凋亡。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-08-19 DOI: 10.1007/s10616-023-00588-w

Lin Cheng, Haoqing Zhai, Juan Du, Gang Zhang, Gan Shi

{"title":"Lobetyolin inhibits cell proliferation and induces cell apoptosis by downregulating ASCT2 in gastric cancer.","authors":"Lin Cheng, Haoqing Zhai, Juan Du, Gang Zhang, Gan Shi","doi":"10.1007/s10616-023-00588-w","DOIUrl":"10.1007/s10616-023-00588-w","url":null,"abstract":"Gastric cancer (GC) is a heterogeneous disease and is the fifth most common cancer worldwide. Lobetyolin, as a bioactive ingredient extracted from Codonopsis pilosula (Franch.) Nannf., has been reported to exert anti-tumor effects in several cancer types. This study was aimed to investigate the role of lobetyolin in GC and the associated mechanism. MKN-45 and MKN-28 cells were incubated with concentrations of lobetyolin for 24 h. The viability and survival of GC cells were evaluated by performing MTT assay. Glutamine uptake, Adenosine Triphosphate, reactive oxygen species (ROS), and glutathione levels were measured by corresponding kits. Apoptosis and mitochondrial membrane potential of GC cells were determined by flow cytometry. Alanine, serine, cysteine-preferring transporter 2 (ASCT2) and the AKT/GSK3β/c-Myc pathway protein levels were examined by western blotting. Xenograft model and immunohistochemical staining were used to evaluate the pharmacological effects of lobetyolin in mice in vivo. We found that lobetyolin treatment suppressed the proliferative capacity of both MKN-45 and MKN-28 cells in a concentration-dependent manner. Lobetyolin reduced the uptake of glutamine and downregulated the expression levels of ASCT2 in GC cells and xenograft tumors. Lobetyolin effectively restrained the growth of tumors in vivo. In addition, lobetyolin induced the accumulation of ROS to attenuate mitochondria-mediated apoptosis via downregulation of ASCT2 expression. Lobetyolin promoted the phosphorylation of c-Myc and suppressed the phosphorylation of GSK3β and AKT in both MKN-45 and MKN-28 cells. The level of total Nrf2 protein was reduced after lobetyolin treatment. Overall, lobetyolin exerts anti-cancer effects by repressing cell proliferation and inducing cell apoptosis via downregulation of ASCT2 in GC.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"435-448"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465467/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137219","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}

引用次数: 0

New high-throughput screening method for Chinese hamster ovary cell lines expressing low reduced monoclonal antibody levels: application of a system controlling the gas phase over cell lysates in miniature bioreactors and facilitating multiple sample setup. 表达低降低单克隆抗体水平的中国仓鼠卵巢细胞系的新高通量筛选方法：在微型生物反应器中控制细胞裂解物气相的系统的应用和促进多样品设置。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-10-01 Epub Date: 2023-07-21 DOI: 10.1007/s10616-023-00587-x

Tsuyoshi Yamaguchi, Mie Fukuda, Yuichi Matsumoto, Takaaki Mori, Shinsuke Kikuchi, Ryuma Nagano, Koichi Yamamoto, Kaori Wakamatsu

{"title":"New high-throughput screening method for Chinese hamster ovary cell lines expressing low reduced monoclonal antibody levels: application of a system controlling the gas phase over cell lysates in miniature bioreactors and facilitating multiple sample setup.","authors":"Tsuyoshi Yamaguchi, Mie Fukuda, Yuichi Matsumoto, Takaaki Mori, Shinsuke Kikuchi, Ryuma Nagano, Koichi Yamamoto, Kaori Wakamatsu","doi":"10.1007/s10616-023-00587-x","DOIUrl":"10.1007/s10616-023-00587-x","url":null,"abstract":"Interchain disulfide bonds in monoclonal antibodies may be reduced during large-scale mAb production using Chinese hamster ovary (CHO) cells. This reaction lowers the mAb product yield and purity; however, it may be prevented by screening cell lines that are unsusceptible to reduction and using them in mAb production. Antibody reduction susceptibility may be cell line-dependent. To the best of our knowledge, however, an efficient method of screening reduction-unsusceptible CHO cell lines has not been previously reported. Here, we report a novel screening method that can simultaneously detect and identify mAb reduction susceptibility in lysates containing ≤ 48 CHO cell lines. This evaluation system was equally effective and generated similar results at all culture scales, including 250 mL, 3 L, and 1000 L. Furthermore, we discovered that reduction-susceptible cell lines contained higher total intracellular nicotinamide adenine dinucleotide phosphate (NADPH) and NADP+ concentrations than reduction-unsusceptible cell lines, regardless of whether they expressed immunoglobulin (Ig)G4 or IgG1. NADPH or NADP+ supplementation in the lysate of reduction-unsusceptible cells resulted in mAb reduction. Application of the innovative CHO cell line screening approach could mitigate or prevent reductions in large-scale mAb generation from CHO cells.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 5","pages":"421-433"},"PeriodicalIF":2.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10465464/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10135186","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}

引用次数: 0

Optimization of adaptation parameters from adhesion cell culture in serum-containing media to suspension in chemically defined media by superlative box design 用最佳箱设计优化含血清培养基中粘附细胞培养到化学定义培养基中悬浮细胞的适应参数

4区生物学

Cytotechnology Pub Date : 2023-09-29 DOI: 10.1007/s10616-023-00596-w

Wanyue Cui, Shijie Liu

引用次数: 0

An agarose-alginate microfluidic device for the study of spheroid invasion, ATRA inhibits CAFs-mediated matrix remodeling. 一种用于研究球体侵袭的琼脂糖-藻酸盐微流体装置，ATRA抑制CAFs介导的基质重塑。

IF 2 4区生物学

Cytotechnology Pub Date : 2023-08-01 Epub Date: 2023-04-11 DOI: 10.1007/s10616-023-00578-y

Mohammad Reza Nasiraee, Shabnam Shahrivari, Soheila Sayad, Hoda Mahdavi, Neda Saraygord-Afshari, Zeinab Bagheri

{"title":"An agarose-alginate microfluidic device for the study of spheroid invasion, ATRA inhibits CAFs-mediated matrix remodeling.","authors":"Mohammad Reza Nasiraee, Shabnam Shahrivari, Soheila Sayad, Hoda Mahdavi, Neda Saraygord-Afshari, Zeinab Bagheri","doi":"10.1007/s10616-023-00578-y","DOIUrl":"10.1007/s10616-023-00578-y","url":null,"abstract":"Growing evidence demonstrates that cancer-associated fibroblasts (CAF) are responsible for tumor genesis, growth, metastasis, and treatment response. Therefore, targeting these cells may contribute to tumor control. It has been proposed that targeting key molecules and pathways of proliferative functions can be more effective than killing CAFs. In this regard, multicellular aggregates, like spheroids, can be used as human tumor models. Spheroids closely resemble human tumors and mimic many of their features. Microfluidic systems are ideal for cultivation and study of spheroids. These systems can be designed with different biological and synthetic matrices in order to have a more realistic simulation of the tumor microenvironment (TME). In this study, we investigated the effect of all-trans retinoic acid (ATRA) on 3D spheroid invasion of MDA-MB cells exposed to hydrogel matrix derived from CAFs. The number of invasive cells significantly decreased in CAF-ECM hydrogel treated with ATRA (p < 0.05), which indicates that ATRA could be effective for CAFs normalization. This experiment was done using an agarose-alginate microfluidic chip. As compared with common methods, such hydrogel casting is an easier method for chip fabrication and can even reduce costs.Graphical abstract: Supplementary information: The online version contains supplementary material available at 10.1007/s10616-023-00578-y.","PeriodicalId":10890,"journal":{"name":"Cytotechnology","volume":"75 4","pages":"309-323"},"PeriodicalIF":2.0,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10299977/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9741988","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}

引用次数: 0