Name: enrichment for chemoresistant ovarian cancer stem cells from human cell lines
Uploaded: 2014-09-10T15:00:00
Description: Watch this Scientific Journal Video about Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines at JoVE.com

Serene Samyesudhas and Dr. Lynn Roy assisted in preparing samples for filming.

1 सेल डिम्बग्रंथि कैंसर कोशिका लाइनों की संस्कृति और फ्लोरोसेंट लेबलिंग <ol><li> SKOV3 मीडिया तैयार: 10% भ्रूण गोजातीय सीरम (FBS) के साथ पूरक मैककॉय मीडिया, 0.1 मिमी एल glutamine, 50 यू / एमएल पेनिसिलिन, और 50 माइक्रोग्राम / एमएल...

<ol>
	<li>Pennington, K., Pulaski, H., Pennington, M., Liu, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Too+Much+of+a+Good+Thing:+Suicide+Prevention+Promotes+Chemoresistance+in+Ovarian+Carcinoma.">Too Much of a Good Thing: Suicide Prevention Promotes Chemoresistance in Ovarian Carcinoma.</a> Curr Cancer Drug Targets. 10, 575-583 (2010).</li><li>Thigpen, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rational+approach+to+the+management+of+recurrent+or+persistent+ovarian+carcinoma.">A rational approach to the management of recurrent or persistent ovarian carcinoma.</a> Clin Obstet Gynecol. 55, 114-130 (2012).</li><li>Burgos-Ojeda, D., Rueda, B. R., Buckanovich, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+cancer+stem+cell+markers:+prognostic+and+therapeutic+implications.">Ovarian cancer stem cell markers: prognostic and therapeutic implications.</a> Cancer letters. 322, 1-7 (2012).</li><li>Ma, L., Lai, D., Liu, T., Cheng, W., Guo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+stem-like+cells+can+be+isolated+with+drug+selection+in+human+ovarian+cancer+cell+line+SKOV3.">Cancer stem-like cells can be isolated with drug selection in human ovarian cancer cell line SKOV3.</a> Acta Biochim Biophys Sin (Shanghai). 42, 593-602 (2010).</li><li>Bonnet, D., Dick, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+acute+myeloid+leukemia+is+organized+as+a+hierarchy+that+originates+from+a+primitive+hematopoietic+cell.">Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell.</a> Nature Medicine. 3, 730-737 (1997).</li><li>Alison, M. R., Lim, S. M., Nicholson, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+stem+cells:+problems+for+therapy.">Cancer stem cells: problems for therapy.</a> The Journal of pathology. 223, 147-161 (2011).</li><li>Luo, X., Dong, Z., Chen, Y., Yang, L., Lai, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enrichment+of+ovarian+cancer+stem-like+cells+is+associated+with+epithelial+to+mesenchymal+transition+through+an+miRNA-activated+AKT+pathway.">Enrichment of ovarian cancer stem-like cells is associated with epithelial to mesenchymal transition through an miRNA-activated AKT pathway.</a> Cell proliferation. 46, 436-446 (2013).</li><li>Wang, L., Mezencev, R., Bowen, N. J., Matyunina, L. V., McDonald, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+stem-like+cells+from+a+human+ovarian+cancer+cell+line.">Isolation and characterization of stem-like cells from a human ovarian cancer cell line.</a> Molecular and cellular biochemistry. 363, 257-268 (2012).</li><li>Abubaker, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+single+treatment+of+chemotherapy+results+in+the+enrichment+of+ovarian+cancer+stem+cell-like+cells+leading+to+an+increased+tumor+burden.">Short-term single treatment of chemotherapy results in the enrichment of ovarian cancer stem cell-like cells leading to an increased tumor burden.</a> Molecular cancer. 12, 24 (2013).</li><li>Conic, I., Dimov, I., Tasic-Dimov, D., Djordjevic, B., Stefanovic, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+epithelial+cancer+stem+cells.">Ovarian epithelial cancer stem cells.</a> ScientificWorldJournal. 11, 1243-1269 (2011).</li><li>Liu, K. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ovarian+cancer+stem-like+cells+show+induced+translineage-differentiation+capacity+and+are+suppressed+by+alkaline+phosphatase+inhibitor.">Ovarian cancer stem-like cells show induced translineage-differentiation capacity and are suppressed by alkaline phosphatase inhibitor.</a> Oncotarget. 4 (12), 2366-2382 (2013).</li><li>Liu, T., Cheng, W., Lai, D., Huang, Y., Guo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+primary+ovarian+cancer+cells+in+different+culture+systems.">Characterization of primary ovarian cancer cells in different culture systems.</a> Oncology reports. 23, 1277-1284 (2010).</li></ol>

उपचार के लिए प्रतिरोधी रहे हैं कि सीएससी प्राथमिक ट्यूमर के इलाज के बाद पतन के लिए जिम्मेदार हो सकता है. सीएससी की विशेषता डिम्बग्रंथि के कैंसर के लिए सुधार के उपचारों के लिए नेतृत्व कर सकते हैं. ऊपर प्?...

Resistance to chemotherapy is a major impediment to the treatment and cure of cancer. Ovarian cancer is the 5th leading cause of cancer-related deaths among women in the United States (American Cancer Society Facts and Figures 2013). Patients initially respond well to chemotherapy, but most patients relapse1. After relapse patients are treated with a variety of additional chemotherapy agents with very little benefit2. General properties of CSCs include unlimited proliferative capabilities, retention of an undifferentiated state, resistance to drug treatment, efficient DNA repair, and ability to generate malignant tumor cells with different phenotypes3. CSCs frequently exhibit expression of stem cell genes such as Nanog, Oct4, Sox2, Nestin, CD133, and CD117. CSCs often express elevated levels of ABCG2, and ALDH genes that may contribute to drug efflux and metabolism3,4. 
The first definitive evidence for CSCs was demonstrated by isolating acute myeloid leukemia-initiating cells that were capable of self-renewal and tumor generation5. These leukemic stem cells expressed surface CD34 and generated leukemia in NOD/SCID (immunocompromised) mice5. Since then CSCs have been identified in many cancer types including leukemias/lymphomas, breast, bladder, colorectal, endometrial, sarcomas, hepatocellular carcinoma, melanomas, gliomas, ovarian, pancreatic, prostate, squamous cell carcinoma, and lung6. Therefore, being able to study this subtype of cancer cell is advantageous.
The goal of this study is to create a protocol for the selection and isolation of chemoresistant CSCs. Several methods have been reported for the isolation of CSCs from ovarian cancer cell lines. Non-adherent spheroids isolated from OVCAR-3, SKOV3, or HO8910 cultures demonstrate stem-like properties7,8. Isolation of CD133+ cells from OVCAR-3 cultures also yields CSCs. CSCs have also been selected in culture by treatment with chemotherapeutic agents. Treating tumor cell lines (OVCA433, Hey, and SKOV3 cells) with cisplatin and paclitaxel allows for the expansion and isolation of CSCs4,9. While culture of some cell types in CSC media leads to isolation of CSCs, SKOV3 cells did not survive culture in serum-free media or form sphere cells4. Therefore, treatment of cells with cisplatin and paclitaxel aided the expansion or isolation of this population4.
Using a modification of the procedure presented by Ma and colleagues4 we developed a method to isolate CSCs from the ovarian cancer cell lines. Our protocol is advantageous as it yields more viable cells while using less toxic chemotherapeutic agents. Cells are treated with cisplatin and subsequently grown in serum-free media supplemented with growth factors (stem cell media). We isolate the resulting non-adherent sphere cells and assay them for their expression of stem cell markers. This model enables the study of CSC properties and response to drug therapy.

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enrichment for chemoresistant ovarian cancer stem cells from human cell lines

कैंसर स्टेम कोशिकाओं (सीएससी) धीमी साइकिल और अत्यधिक, proliferative आक्रामक उत्पन्न करने के लिए asymmetrically कि विभाजन undifferentiated कोशिकाओं, और chemoresistant ट्यूमर कोशिकाओं के एक सबसेट के रूप में परिभाषित कर रहे हैं. इसलिए, सीएससी therapeutically लक्षित करने के लिए कोशिकाओं का एक आकर्षक आबादी हैं. सीएससी रक्त, मस्तिष्क, फेफड़े, जठरांत्र संबंधी मार्ग, प्रोस्टेट, और अंडाशय में उन सहित कैंसर के प्रकार के एक नंबर के लिए योगदान करने के लिए भविष्यवाणी कर रहे हैं. अलग और सीएससी के लिए एक ट्यूमर सेल आबादी सीएससी के गुण, जेनेटिक्स, और चिकित्सीय प्रतिक्रिया का अध्ययन करने के लिए सक्षम शोधकर्ताओं जाएगा समृद्ध. हम reproducibly डिम्बग्रंथि के कैंसर कोशिका लाइनों (SKOV3 और OVCA429) से डिम्बग्रंथि के कैंसर सीएससी के लिए समृद्ध करती है कि एक प्रोटोकॉल उत्पन्न. सेल लाइनों 3 दिन के लिए 20 माइक्रोन सिसप्लैटिन के साथ व्यवहार कर रहे हैं. जीवित कोशिकाओं साइटोकिन्स और वृद्धि कारकों युक्त सीरम मुक्त स्टेम सेल मीडिया में पृथक और सुसंस्कृत हैं. हम कश्मीर के लिए अलग कक्षों का विश्लेषण करके इन शुद्ध सीएससी की एक संवर्धन का प्रदर्शनNown सेल मार्कर Oct4, Nanog, और Prom1 (CD133) और CD177 और CD133 की कोशिका की सतह अभिव्यक्ति स्टेम. सीएससी प्रदर्शनी chemoresistance वृद्धि हुई. सीएससी के अलगाव के लिए इस विधि chemoresistance और ट्यूमर पतन में सीएससी की भूमिका के अध्ययन के लिए एक उपयोगी उपकरण है.

हम सिसप्लैटिन उपचार का उपयोग उपकला डिम्बग्रंथि के कैंसर कोशिका लाइनों से सीएससी पृथक दिखाना है कि, हम पहले पूर्व उपचार के लिए और चयन के बाद सेल लाइनों की छवियों का अधिग्रहण किया. हम पक्षपाती (इलाज) SKOV3 और ...

Watch this Scientific Journal Video about Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines at JoVE.com

Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines

कैंसर स्टेम कोशिकाओं (सीएससी) के कारण chemoresistance को ट्यूमर पतन में फंसा रहे हैं. हम चयन गर्भाशय के कैंसर कोशिका लाइनों से सीएससी के विस्तार के लिए एक प्रोटोकॉल अनुकूलित है. Chemotherapeutic सिसप्लैटिन साथ कोशिकाओं के इलाज और हम गैर पक्षपाती सीएससी संस्कृतियों के लिए बेहतर मीडिया को बढ़ावा देने के लिए एक स्टेम सेल में संवर्धन द्वारा.

मानव कोशिका लाइनों से Chemoresistant डिम्बग्रंथि के कैंसर स्टेम सेल के लिए संवर्धन

Cancer stem cells (CSCs) are defined as a subset of slow cycling and undifferentiated cells that divide asymmetrically to generate highly proliferative, ...

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Brain tumors are typically comprised of morphologically diverse cells that express a variety of neural lineage markers. Only a relatively small fraction ...

Isolation of Cardiomyocyte Nuclei from Post-mortem Tissue

Cardiomyocyte नाभिक का पोस्टमार्टम ऊतक से अलगाव

Identification of cardiomyocyte nuclei has been challenging in tissue sections as most strategies rely only on cytoplasmic marker proteins1. Rare events ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

मानव मानव neuroendocrine ट्यूमर चिकित्सा के अध्ययन के लिए एक तीन आयामी मॉडल के रूप में neuroendocrine ट्यूमर सेल लाइनों

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

मानव दंत लुगदी स्टेम सेल के अलगाव विशेषता, और तुलनात्मक भेदभाव दो अलग अलग तरीकों का उपयोग करके स्थायी दांत से व्युत्पन्न

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

Time-lapse Imaging of Primary Preneoplastic Mammary Epithelial Cells Derived from Genetically Engineered Mouse Models of Breast Cancer

प्राथमिक Preneoplastic स्तन उपकला कोशिकाओं के समय चूक इमेजिंग स्तन कैंसर के आनुवंशिक रूप से इंजीनियर माउस मॉडल से निकाली

Time-lapse  imaging can be used to compare behavior of cultured primary preneoplastic  mammary epithelial cells derived from different genetically ...

Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma

Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma

रोगी व्युत्पन्न सेल संस्कृति और CD133 का अलगाव<sup&gt; +</sup&gt; मेलेनोमा से ख्यात कैंसर स्टेम सेल

Despite improved treatments options for melanoma available today, patients with advanced malignant melanoma still have a poor prognosis for ...

Isolation of Cancer Stem Cells From Human Prostate Cancer Samples

मानव प्रोस्टेट कैंसर के नमूनों से कैंसर स्टेम कोशिकाओं के अलगाव

The cancer stem cell (CSC) model has been considerably revisited over the last two decades. During this time CSCs have been identified and directly ...

Method for Obtaining Primary Ovarian Cancer Cells From Solid Specimens

ठोस नमूनों से प्राथमिक डिम्बग्रंथि कैंसर कोशिकाओं को प्राप्त करने के लिए विधि

Reliable tools for investigating ovarian cancer initiation and progression are urgently needed. While the use of ovarian cancer cell lines remains a ...

Evaluation of Stem Cell Properties in Human Ovarian Carcinoma Cells Using Multi and Single Cell-based Spheres Assays

मल्टी और सिंगल सेल आधारित क्षेत्रों assays का उपयोग मानव डिम्बग्रंथि कार्सिनोमा कोशिकाओं में स्टेम सेल गुणों का मूल्यांकन

Years of research indicates that ovarian cancers harbor a heterogeneous mixture of cells including a subpopulation of so-called &#8220;cancer stem ...

In vitro Enrichment of Ovarian Cancer Tumor-initiating Cells

In vitro Enrichment of Ovarian Cancer Tumor-initiating Cells

डिम्बग्रंथि के कैंसर के ट्यूमर शुरुआत कोशिकाओं की इन विट्रो संवर्धन में

Evidence suggests that small subpopulations of tumor cells maintain a unique self-renewing and differentiation capacity and may be responsible for tumor ...