Cell biology international reports最新文献_第3页

Selective silencing of DNA topoisomerase IIβ in human mesenchymal stem cells by siRNAs (small interfering RNAs) 小干扰rna对人间充质干细胞DNA拓扑异构酶ⅱβ的选择性沉默

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20110003

Nebiyyeh Kamaci, Tuba Emnacar, Nihal Karakas, Gulsum Arikan, Ken Tsutsui, Sevim Isik

{"title":"Selective silencing of DNA topoisomerase IIβ in human mesenchymal stem cells by siRNAs (small interfering RNAs)","authors":"Nebiyyeh Kamaci, Tuba Emnacar, Nihal Karakas, Gulsum Arikan, Ken Tsutsui, Sevim Isik","doi":"10.1042/CBR20110003","DOIUrl":"10.1042/CBR20110003","url":null,"abstract":"hMSCs (human mesenchymal stem cells) express two isoforms of DNA topo II (topoisomerase II). Although both isoforms have the same catalytic activity, they are specialized for different functions in the cell: while topo IIα is essential for chromosome segregation in mitotic cells, topo IIβ is involved in more specific cellular functions. A number of inhibitors are available that inhibit the catalytic activity of both topo II isoforms. However, in order to investigate the isoform-specific inhibition of these two enzymes, it is necessary to use other techniques such as siRNA (small interfering RNA) interference to selectively silence either one of the isoforms individually. Depending on the lipid charge densities and protein varieties of the cell membrane, previous studies have demonstrated that transfection efficiencies of siRNAs to hMSCs are very low. In the study reported here, we demonstrate the use of Lipofectamine RNAiMAX as an efficient transfection reagent to introduce siRNAs into human mesenchymal stem cells with significantly great efficiency to silence topo IIβ selectively. A high level of transfection efficiency (80%) was achieved by using unlabelled topo IIβ-specific siRNA oligos. Specifically, it was confirmed repeatedly that green labelled siRNAs interfere with the transfection of siRNAs. The reagent induced minimal cytotoxicity (3.5–4.5%), and cell viability of the transfected hMSCs decreased 20–30% compared with untreated cells, depending on the concentration of the reagent.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"18 1","pages":"15-21"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20110003","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31020784","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

Establishment of glass catfish (Kryptopterus bicirrhis) fin-derived cells 玻璃鲶鱼鱼鳍细胞的建立

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20110002

Jee Eun Han, Casiano H Choresca, Ok Jae Koo, Hyun Ju Oh, So Gun Hong, Ji Hyung Kim, Sang Phil Shin, Jin Woo Jun, Byeong Chun Lee, Se Chang Park

{"title":"Establishment of glass catfish (Kryptopterus bicirrhis) fin-derived cells","authors":"Jee Eun Han, Casiano H Choresca, Ok Jae Koo, Hyun Ju Oh, So Gun Hong, Ji Hyung Kim, Sang Phil Shin, Jin Woo Jun, Byeong Chun Lee, Se Chang Park","doi":"10.1042/CBR20110002","DOIUrl":"10.1042/CBR20110002","url":null,"abstract":"Genetically manipulated transparent animals were already explored in many species for in vivo study of gene expression, transplantation analysis and cancer biology. However, there are no reports about transparent animals as in vitro genetic resources. In the present study, fin-derived cells from glass catfish (Krytopterus bicirrhis), naturally transparent fish with a visible skeleton and internal organs, were isolated after culturing fin explants and characterized using cryopreservation and cell cycle analysis. The cells grew well in DMEM (Dulbecco's modified Eagle's medium) containing 1% (v/v) P/S (penicillin—streptomycin) and 10% (v/v) fetal bovine serum at 26°C and showed increased cryopreservation efficiency with the slow-freezing method in the presence of 15% dimethyl sulfoxide. In addition, cell cycle analysis was evaluated based on flow cytometric analysis, and culturing to confluence (>85%) was more effective for synchronizing cells at the G0/G1 stages than roscovitine treatment (<75%). This is the first report about cell isolation from transparent animals. The results from testing the cell's viability following cryopreservation and subjecting the cells to cycle analysis can be useful tools for genetic resource management.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"18 1","pages":"1-5"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20110002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31020783","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 12

hMSCs possess the potential to differentiate into DP cells in vivo and in vitro hMSCs在体内和体外均具有向DP细胞分化的潜力

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20120003

Minjuan Wu, Qing Sun, Xiaocan Guo, Houqi Liu

引用次数: 19

Identification of functional tissue-resident cardiac stem/progenitor cells in adult mouse 成年小鼠功能性组织驻留心脏干/祖细胞的鉴定

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20120001

Leilei Lu, Fusheng Li, Jingjing Lu

{"title":"Identification of functional tissue-resident cardiac stem/progenitor cells in adult mouse","authors":"Leilei Lu, Fusheng Li, Jingjing Lu","doi":"10.1042/CBR20120001","DOIUrl":"10.1042/CBR20120001","url":null,"abstract":"In most somatic tissues, ASCs (adult stem cells) are crucial for the maintenance of tissue homoeostasis under normal physiological state and recovery from injury. LRC (label retaining cell) assay is a well-known method of identifying possible somatic stem/progenitor cells and their location both in situ and in vivo. BrdU (bromodeoxyuridine) was used here to tag the possible CSCs (cardiac stem cells)/CPCs (cardiac progenitor cells) in newborn pups, followed by a trace period of up to 24 months. In addition, we have used our newly developed ‘KAL’ method to rapidly Kill proliferating cells in adult heart tissues, then, Activate and Label the surviving CSCs/CPCs. LRCs that definitively exist in the heart tissues of adult mice, and some LRCs express the stem cell marker, Sca-1 or c-Kit, and are located primarily in the myocardium and vascular endothelial regions. Moreover, the number of LRCs remains nearly constant during the lifespan of the mouse. After injury induced by 5-fluorouracil, the proliferating cells were almost completely cleared on day 3, and the activated CSCs/CPCs retained their BrdU label after regeneration was complete. A small percentage of the CSCs/CPCs express Sca-1 or c-Kit. Furthermore, the LRC method together with KAL may be used to identify and locate possible CSCs/CPCs, which has potential clinical application.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"19 1","pages":"15-22"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20120001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31026130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Antigenotoxic and antioxidant activity in human chronic myelogenous leukaemia cell line K562 enhanced by Nitraria retusa leaf extracts 白刺叶提取物增强人慢性髓性白血病K562细胞的抗氧化活性

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1002/cbi3.10003

Jihed Boubaker, Zied Ghedira, Kamel Ghedira, Leila Chekir-Ghedira

引用次数: 2

Human endometrial stem cells as a new source for programming to neural cells 人类子宫内膜干细胞作为神经细胞编程的新来源

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20110009

Zahra Taherian Mobarakeh, Jafar Ai, Farzad Yazdani, Seyed Mahdi Rezayat Sorkhabadi, Zinat Ghanbari, Abbas Noroozi Javidan, Seyed Abdol Reza Mortazavi-Tabatabaei, Mohammad Massumi, Somayeh Ebrahimi Barough

{"title":"Human endometrial stem cells as a new source for programming to neural cells","authors":"Zahra Taherian Mobarakeh, Jafar Ai, Farzad Yazdani, Seyed Mahdi Rezayat Sorkhabadi, Zinat Ghanbari, Abbas Noroozi Javidan, Seyed Abdol Reza Mortazavi-Tabatabaei, Mohammad Massumi, Somayeh Ebrahimi Barough","doi":"10.1042/CBR20110009","DOIUrl":"10.1042/CBR20110009","url":null,"abstract":"Human EnSC (endometrial-derived stem cell) is an abundant and easily available source for cell replacement therapy. Many investigations have shown the potency of the cells to differentiate into several mesoderm-derived cell lineages, including osteocytes and adipocytes. Here, the potency of EnSC in neural differentiation has been investigated. Flow cytometric analysis showed that they were positive for CD90, CD105, OCT4, CD44 and negative for CD31, CD34, CD133. The characterized cells were induced into neural differentiation by bFGF (basic fibroblast growth factor), PDGF (platelet-derived growth factor) and EGF (epidermal growth factor) signalling molecules, respectively in a sequential protocol, and differentiated cells were analysed for expression of neuronal markers by RT—PCR (reverse transcription—PCR) and immunocytochemistry, including Nestin, GABA (γ-aminobutyric acid), MAP2 (microtubule-associated protein 2), β3-tub (class III β-tubulin) and NF-L (neurofilament-light) at the level of their mRNAs. The expression of MAP2, β3-tub and NF-L proteins in EnSC was confirmed 28 days PT (post-treatment) by immunocytochemistry. In conclusion, EnSC can respond to signalling molecules that are usually used as standards in neural differentiation and can programme neuronal cells, making these cells worth considering as a unique source for cell therapy in neurodegenerative disease.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"19 1","pages":"7-14"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20110009","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31025299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 60

Etoposide sensitizes neuroblastoma cells expressing caspase 8 to TRAIL 依托泊苷使表达caspase 8的神经母细胞瘤细胞对TRAIL增敏

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20110008

Hye Ryung Kim, Myoung Woo Lee, Dae Seong Kim, Ha Yeong Jo, Soo Hyun Lee, Hee Won Chueh, Hye Lim Jung, Keon Hee Yoo, Ki Woong Sung, Hong Hoe Koo

{"title":"Etoposide sensitizes neuroblastoma cells expressing caspase 8 to TRAIL","authors":"Hye Ryung Kim, Myoung Woo Lee, Dae Seong Kim, Ha Yeong Jo, Soo Hyun Lee, Hee Won Chueh, Hye Lim Jung, Keon Hee Yoo, Ki Woong Sung, Hong Hoe Koo","doi":"10.1042/CBR20110008","DOIUrl":"10.1042/CBR20110008","url":null,"abstract":"TRAIL [TNF (tumour necrosis factor)-related apoptosis-inducing ligand] is a promising agent for clinical use since it kills a wide range of tumour cells without affecting normal cells. We provide evidence that pretreatment with etoposide significantly enhanced TRAIL-mediated apoptosis via up-regulation of DR5 (death receptor 5 or TRAIL-R2) expression in the caspase 8 expressing neuroblastoma cell line, SK-N-MC. In addition, sequential treatment with etoposide and TRAIL increased caspases 8, 9 and 3 activation, Mcl-1 cleavage and Bid truncation, which suggests that the ability of etoposide and TRAIL to induce apoptosis is mediated through activation of an intrinsic signalling pathway. Although TRAIL-R2 expression increased in IMR-32 cells in response to etoposide treatment, cell death was not increased by concurrent treatment with TRAIL compared with etoposide alone, because the cells lacked caspase 8 expression. Restoration of caspase 8 expression by exposure to IFNγ (interferon γ) sensitizes IMR-32 cells to TRAIL. Moreover, pretreatment with etoposide increased TRAIL-induced apoptosis in caspase 8 restored IMR-32 cells through activation of a caspase cascade that included caspases 8, 9 and 3. These results indicate that the etoposide-mediated sensitization of neuroblastoma cells to TRAIL is associated with an increase in TRAIL-R2 expression and requires caspase 8 expression. These observations support the potential use of a combination of etoposide and TRAIL in future clinical trials.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"19 1","pages":"23-30"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20110008","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"31026677","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7

Comparative proteomics of skeletal muscle mitochondria from myostatin-null mice 肌生成抑制素缺失小鼠骨骼肌线粒体的比较蛋白质组学研究

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20110006

Jonathan Puddick, Ryan D. Martinus

引用次数: 7

High-quality RNA extraction from rat pancreatic islet 大鼠胰岛高质量RNA的提取

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1002/cbi3.10002

Takayoshi Kiba, Masahiro Tanemura, Kiyomi Yagyu

{"title":"High-quality RNA extraction from rat pancreatic islet","authors":"Takayoshi Kiba, Masahiro Tanemura, Kiyomi Yagyu","doi":"10.1002/cbi3.10002","DOIUrl":"10.1002/cbi3.10002","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 In recent years, increasing interest surrounding islet replacement therapies in human has provided the drive for advances in the methods used to isolate from humans as well as a host of animal research models. However, there has been no reports describing a technique that reliably improves the quality of RNA extracted from rat pancreatic islet. Male Sprague–Dawley rats, 10- to 12-week-old, were housed in a certified animal care facility. The rats were underwent bile duct cannulation with pancreatic inflation after clamping the distal common bile duct. The pancreas was excised and digested with ETK/Liberase TL solution at 37°C for 30 min without shaking. Cold ETK was added to stop digestion. The islets were purified by discontinuous iodixanol density gradients of 25, 23, 20 and 11% in a modified ETK/OptiPrep® solution. After a 15 min centrifugation at 1,000g, islets were collected from the interface between the 20 and 11% layer. Immediately after purification, islets were used for RNA extraction. RNA was extracted from isolated rat pancreatic islet cells, using the commercially available kit. In the present study, we have described a technique that reliably improves the quality of RNA extracted from rat pancreatic islet using the perfusion technique in the bile duct. The islet cells that we isolated using this technique were suitable for high quality RNA extraction.\u0000 </section>\u0000 </div>","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"20 1","pages":"1-4"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cbi3.10002","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"51263701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 9

Micronucleus formation, proliferative status, cell death and DNA damage in ethosuximide-treated human lymphocytes 乙磺酰亚胺处理的人淋巴细胞微核形成、增殖状态、细胞死亡和DNA损伤

Cell biology international reports Pub Date : 2013-06-25 DOI: 10.1042/CBR20100007

Flávia G Ghiraldini, Maria Luiza S Mello

{"title":"Micronucleus formation, proliferative status, cell death and DNA damage in ethosuximide-treated human lymphocytes","authors":"Flávia G Ghiraldini, Maria Luiza S Mello","doi":"10.1042/CBR20100007","DOIUrl":"10.1042/CBR20100007","url":null,"abstract":"Ethosuximide is a T-type Ca2+ channel blocker that has been used as an anticonvulsant to treat absence seizures. Because no data have yet been reported on the cytotoxicity, genotoxicity and cell death effects of this drug, we have investigated ethosuximide-treated human lymphocytes with Fenech's CBMN (cytochalasin-blocked micronucleus cytome) assay and single-cell gel electrophoresis (comet assay). The tests allowed us to examine micronucleus formation, the proliferative status of the viable cells, nuclear blebbing, nucleoplasmic bridge formation, cell death (CBMN assay) and DNA damage (comet assay). The lymphocytes were treated for 24 h with 25, 50 or 100 μg/ml ethosuximide. The cells used for the CBMN assay were examined either immediately or 24 h after the cytochalasin blockade. For the comet assay, cells were examined immediately or 22 h after 2 h ethosuximide treatment. The results indicate that 25 and 50 μg/ml ethosuximide did not induce micronucleus formation, nuclear abnormalities, cell death or DNA damage, nor did they affect cell proliferation, suggesting no cytotoxic or genotoxic effects under such experimental conditions. However, under treatment with 100 μg/ml ethosuximide, an increase in micronucleus formation and nuclear abnormalities, but a decrease in cell proliferation and in DNA damage, and no change in the cell death ratio, were detected. Although apparently contradictory, the data obtained with the 100 μg/ml concentration may indicate that induction of cytotoxicity and genotoxicity are not to be disregarded when considering this drug concentration. The mechanisms underlying the cellular response to ethosuximide remain to be explored.","PeriodicalId":75683,"journal":{"name":"Cell biology international reports","volume":"17 1","pages":"27-31"},"PeriodicalIF":0.0,"publicationDate":"2013-06-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1042/CBR20100007","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"58439533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 6