Name: dual effects of melanoma cell-derived factors on bone marrow adipocytes differentiation
Uploaded: 2018-08-23T12:30:00
Description: Watch this Scientific Journal Video about Dual Effects of Melanoma Cell-derived Factors on Bone Marrow Adipocytes Differentiation at JoVE.com

Мы благодарим дов Zipori (Институт Вейцмана, Реховот, Израиль) любезно за предоставление нам стромальных клеток костного мозга мышиных линия 14F1.1. Это исследование было поддержано грантов от китайского национального фонда естественных наук (№ 81771729) и Yongchuan больница Чунцин медицинского университета (Nos. YJQN201330; YJZQN201527).

Примечание: Все клетки, используемые в настоящем протоколе должна быть выращены для по крайней мере трех поколений после оттаивания из замороженных запасов клеток.
1. урожай факторы клеток меланомы
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Подготовка
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<li>Получите B16F10 клеток и линии клетки мелан?...

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	<li>Wang, J., 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=Adipogenic+niches+for+melanoma+cell+colonization+and+growth+in+bone+marrow.">Adipogenic niches for melanoma cell colonization and growth in bone marrow.</a> Laboratory Investigation. 97 (6), 737-745 (2017).</li><li>Trotter, T. N., 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=Adipocyte-Lineage+Cells+Support+Growth+and+Dissemination+of+Multiple+Myeloma+in+Bone.">Adipocyte-Lineage Cells Support Growth and Dissemination of Multiple Myeloma in Bone.</a> The American Journal of Pathology. 186 (11), 3054-3063 (2016).</li><li>Morris, E. V., Edwards, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+bone+marrow+adipocytes+in+bone+metastasis.">The role of bone marrow adipocytes in bone metastasis.</a> Journal of Bone Oncology. 5 (3), 121-123 (2016).</li><li>Diedrich, J. D., 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=Bone+marrow+adipocytes+promote+the+Warburg+phenotype+in+metastatic+prostate+tumors+via+HIF-1alpha+activation.">Bone marrow adipocytes promote the Warburg phenotype in metastatic prostate tumors via HIF-1alpha activation.</a> Oncotarget. 7 (40), 64854-64877 (2016).</li><li>Chkourko Gusky, H., Diedrich, J., MacDougald, O. A., Podgorski, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Omentum+and+bone+marrow:+how+adipocyte-rich+organs+create+tumour+microenvironments+conducive+for+metastatic+progression.">Omentum and bone marrow: how adipocyte-rich organs create tumour microenvironments conducive for metastatic progression.</a> Obesity Reviews. 17 (11), 1015-1029 (2016).</li><li>Chen, G. L., 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=High+fat+diet+increases+melanoma+cell+growth+in+the+bone+marrow+by+inducing+osteopontin+and+interleukin+6.">High fat diet increases melanoma cell growth in the bone marrow by inducing osteopontin and interleukin 6.</a> Oncotarget. 7 (18), 26653-26669 (2016).</li><li>Balaban, S., 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=Adipocyte+lipolysis+links+obesity+to+breast+cancer+growth:+adipocyte-derived+fatty+acids+drive+breast+cancer+cell+proliferation+and+migration.">Adipocyte lipolysis links obesity to breast cancer growth: adipocyte-derived fatty acids drive breast cancer cell proliferation and migration.</a> Cancer &amp; Metabolism. 5, 1 (2017).</li><li>Huang, C. 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=Adipocytes+promote+malignant+growth+of+breast+tumours+with+monocarboxylate+transporter+2+expression+via+beta-hydroxybutyrate.">Adipocytes promote malignant growth of breast tumours with monocarboxylate transporter 2 expression via beta-hydroxybutyrate.</a> Nature Communications. 8, 14706 (2017).</li><li>Wang, Y. Y., 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=Mammary+adipocytes+stimulate+breast+cancer+invasion+through+metabolic+remodeling+of+tumor+cells.">Mammary adipocytes stimulate breast cancer invasion through metabolic remodeling of tumor cells.</a> JCI Insight. 2 (4), 87489 (2017).</li><li>Wang, C., Gao, C., Meng, K., Qiao, H., Wang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+adipocytes+stimulate+invasion+of+breast+cancer+MCF-7+cells+by+secreting+IGFBP-2.">Human adipocytes stimulate invasion of breast cancer MCF-7 cells by secreting IGFBP-2.</a> PLoS One. 10 (3), 0119348 (2015).</li><li>Nieman, K. M., 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=Adipocytes+promote+ovarian+cancer+metastasis+and+provide+energy+for+rapid+tumor+growth.">Adipocytes promote ovarian cancer metastasis and provide energy for rapid tumor growth.</a> Nature Medicine. 17 (11), 1498-1503 (2011).</li><li>Herroon, M. 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=Bone+marrow+adipocytes+promote+tumor+growth+in+bone+via+FABP4-dependent+mechanisms.">Bone marrow adipocytes promote tumor growth in bone via FABP4-dependent mechanisms.</a> Oncotarget. 4 (11), 2108-2123 (2013).</li><li>Tabe, Y., 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=Bone+Marrow+Adipocytes+Facilitate+Fatty+Acid+Oxidation+Activating+AMPK+and+a+Transcriptional+Network+Supporting+Survival+of+Acute+Monocytic+Leukemia+Cells.">Bone Marrow Adipocytes Facilitate Fatty Acid Oxidation Activating AMPK and a Transcriptional Network Supporting Survival of Acute Monocytic Leukemia Cells.</a> Cancer Research. 77 (6), 1453-1464 (2017).</li><li>Wen, Y. A., 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=Adipocytes+activate+mitochondrial+fatty+acid+oxidation+and+autophagy+to+promote+tumor+growth+in+colon+cancer.">Adipocytes activate mitochondrial fatty acid oxidation and autophagy to promote tumor growth in colon cancer.</a> Cell Death &amp; Differentiation. 8 (2), 2593 (2017).</li><li>Liu, Z., 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=Mature+adipocytes+in+bone+marrow+protect+myeloma+cells+against+chemotherapy+through+autophagy+activation.">Mature adipocytes in bone marrow protect myeloma cells against chemotherapy through autophagy activation.</a> Oncotarget. 6 (33), 34329-34341 (2015).</li><li>Ye, H., 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=Leukemic+Stem+Cells+Evade+Chemotherapy+by+Metabolic+Adaptation+to+an+Adipose+Tissue+Niche.">Leukemic Stem Cells Evade Chemotherapy by Metabolic Adaptation to an Adipose Tissue Niche.</a> Cell Stem Cell. 19 (1), 23-37 (2016).</li><li>Templeton, Z. S., 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=Breast+Cancer+Cell+Colonization+of+the+Human+Bone+Marrow+Adipose+Tissue+Niche.">Breast Cancer Cell Colonization of the Human Bone Marrow Adipose Tissue Niche.</a> Neoplasia. 17 (12), 849-861 (2015).</li><li>Daquinag, A. C., Souza, G. R., Kolonin, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose+tissue+engineering+in+three-dimensional+levitation+tissue+culture+system+based+on+magnetic+nanoparticles.">Adipose tissue engineering in three-dimensional levitation tissue culture system based on magnetic nanoparticles.</a> Tissue Engineering Part C: Methods. 19 (5), 336-344 (2013).</li><li>Emont, M. P., 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=Using+a+3D+Culture+System+to+Differentiate+Visceral+Adipocytes+In+Vitro.">Using a 3D Culture System to Differentiate Visceral Adipocytes In Vitro.</a> Endocrinology. 156 (12), 4761-4768 (2015).</li><li>Katt, M. E., Placone, A. L., Wong, A. D., Xu, Z. S., Searson, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+Vitro+Tumor+Models:+Advantages,+Disadvantages,+Variables,+and+Selecting+the+Right+Platform.">In Vitro Tumor Models: Advantages, Disadvantages, Variables, and Selecting the Right Platform.</a> Frontiers in Bioengineering and Biotechnology. 4, 12 (2016).</li><li>Edmondson, R., Broglie, J. J., Adcock, A. F., Yang, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+cell+culture+systems+and+their+applications+in+drug+discovery+and+cell-based+biosensors.">Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors.</a> ASSAY and Drug Development Technologies. 12 (4), 207-218 (2014).</li><li>Herroon, M. K., Diedrich, J. D., Podgorski, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+3D-Culture+Approaches+to+Study+Interactions+of+Bone+Marrow+Adipocytes+with+Metastatic+Prostate+Cancer+Cells.">New 3D-Culture Approaches to Study Interactions of Bone Marrow Adipocytes with Metastatic Prostate Cancer Cells.</a> Frontiers in Endocrinology (Lausanne). 7, 84 (2016).</li><li>Lee, J. M., 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=A+three-dimensional+microenvironment+alters+protein+expression+and+chemosensitivity+of+epithelial+ovarian+cancer+cells+in+vitro.">A three-dimensional microenvironment alters protein expression and chemosensitivity of epithelial ovarian cancer cells in vitro.</a> Laboratory Investigation. 93 (5), 528-542 (2013).</li><li>Imamura, Y., 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=Comparison+of+2D-+and+3D-culture+models+as+drug-testing+platforms+in+breast+cancer.">Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer.</a> Oncology Reports. 33 (4), 1837-1843 (2015).</li><li>Birgersdotter, A., Sandberg, R., Ernberg, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+expression+perturbation+in+vitro--a+growing+case+for+three-dimensional+(3D)+culture+systems.">Gene expression perturbation in vitro--a growing case for three-dimensional (3D) culture systems.</a> Seminars in Cancer Biology. 15 (5), 405-412 (2005).</li><li>Wang, W., 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=3D+spheroid+culture+system+on+micropatterned+substrates+for+improved+differentiation+efficiency+of+multipotent+mesenchymal+stem+cells.">3D spheroid culture system on micropatterned substrates for improved differentiation efficiency of multipotent mesenchymal stem cells.</a> Biomaterials. 30 (14), 2705-2715 (2009).</li><li>Dirat, B., 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=Cancer-associated+adipocytes+exhibit+an+activated+phenotype+and+contribute+to+breast+cancer+invasion.">Cancer-associated adipocytes exhibit an activated phenotype and contribute to breast cancer invasion.</a> Cancer Research. 71 (7), 2455-2465 (2011).</li><li>Scott, M. A., Nguyen, V. T., Levi, B., James, A. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+methods+of+adipogenic+differentiation+of+mesenchymal+stem+cells.">Current methods of adipogenic differentiation of mesenchymal stem cells.</a> Stem Cells and Development. 20 (10), 1793-1804 (2011).</li><li>Zipori, D., Toledo, J., von der Mark, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+heterogeneity+among+stromal+cell+lines+from+mouse+bone+marrow+disclosed+in+their+extracellular+matrix+composition+and+interactions+with+normal+and+leukemic+cells.">Phenotypic heterogeneity among stromal cell lines from mouse bone marrow disclosed in their extracellular matrix composition and interactions with normal and leukemic cells.</a> Blood. 66 (2), 447-455 (1985).</li><li>Maridas, D. E., Rendina-Ruedy, E., Le, P. T., Rosen, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation,+Culture,+and+Differentiation+of+Bone+Marrow+Stromal+Cells+and+Osteoclast+Progenitors+from+Mice.">Isolation, Culture, and Differentiation of Bone Marrow Stromal Cells and Osteoclast Progenitors from Mice.</a> Journal of Visualized Experiments. (131), e56750 (2018).</li><li>Iguchi, T., Niino, N., Tamai, S., Sakurai, K., Mori, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Absolute+Quantification+of+Plasma+MicroRNA+Levels+in+Cynomolgus+Monkeys,+Using+Quantitative+Real-time+Reverse+Transcription+PCR.">Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR.</a> Journal of Visualized Experiments. (132), e56850 (2018).</li><li>Bozec, A., Hannemann, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+Regulation+of+Adipocyte+Numbers+in+Adult+Organisms+Through+Differentiation+and+Apoptosis+Homeostasis.">Mechanism of Regulation of Adipocyte Numbers in Adult Organisms Through Differentiation and Apoptosis Homeostasis.</a> Journal of Visualized Experiments. (112), e53822 (2016).</li><li>Shafat, M. S., 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=Leukemic+blasts+program+bone+marrow+adipocytes+to+generate+a+protumoral+microenvironment.">Leukemic blasts program bone marrow adipocytes to generate a protumoral microenvironment.</a> Blood. 129 (10), 1320-1332 (2017).</li><li>Gazi, E., 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=Direct+evidence+of+lipid+translocation+between+adipocytes+and+prostate+cancer+cells+with+imaging+FTIR+microspectroscopy.">Direct evidence of lipid translocation between adipocytes and prostate cancer cells with imaging FTIR microspectroscopy.</a> The Journal of Lipid Research. 48 (8), 1846-1856 (2007).</li><li>Brown, M. D., 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=Influence+of+omega-6+PUFA+arachidonic+acid+and+bone+marrow+adipocytes+on+metastatic+spread+from+prostate+cancer.">Influence of omega-6 PUFA arachidonic acid and bone marrow adipocytes on metastatic spread from prostate cancer.</a> British Journal of Cancer. 102 (2), 403-413 (2010).</li><li>Hardaway, A. L., Herroon, M. K., Rajagurubandara, E., Podgorski, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Marrow+adipocyte-derived+CXCL1+and+CXCL2+contribute+to+osteolysis+in+metastatic+prostate+cancer.">Marrow adipocyte-derived CXCL1 and CXCL2 contribute to osteolysis in metastatic prostate cancer.</a> Clinical &amp; Experimental Metastasis. 32 (4), 353-368 (2015).</li><li>Aebi, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+metastasis+in+the+elderly.">Spinal metastasis in the elderly.</a> European Spine Journal. 12, 202-213 (2003).</li><li>Wagner, M., Bjerkvig, R., Wiig, H., Dudley, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Loss+of+adipocyte+specification+and+necrosis+augment+tumor-associated+inflammation.">Loss of adipocyte specification and necrosis augment tumor-associated inflammation.</a> Adipocyte. 2 (3), 176-183 (2013).</li><li>Bochet, L., 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=Cancer-associated+adipocytes+promotes+breast+tumor+radioresistance.">Cancer-associated adipocytes promotes breast tumor radioresistance.</a> Biochemical and Biophysical Research Communications. 411 (1), 102-106 (2011).</li><li>Hirano, T., 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=Enhancement+of+adipogenesis+induction+by+conditioned+media+obtained+from+cancer+cells.">Enhancement of adipogenesis induction by conditioned media obtained from cancer cells.</a> Cancer Letters. 268 (2), 286-294 (2008).</li><li>Gordeev, A. A., Chetverina, H. V., Chetverin, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Planar+arrangement+of+eukaryotic+cells+in+merged+hydrogels+combines+the+advantages+of+3-D+and+2-D+cultures.">Planar arrangement of eukaryotic cells in merged hydrogels combines the advantages of 3-D and 2-D cultures.</a> Biotechniques. 52 (5), 325-331 (2012).</li></ol>

Cocultures с вставками широко использовался для изучения взаимодействия к ячейке. 2D coculture система является эффективным способом наблюдать, как две части перекрестных помех в пробирке, в которой мы здесь показали два различных раковых клеток driven воздействия на адипоциты костного мозга....

Метастазы в кости широко распространены среди больных раком, но курс лечения по-прежнему недоступна. Помимо специализируется на хранение энергии в виде жира, адипоциты может поддерживать рост опухоли и метастазов в костный мозг и другие органы1,2,3,4,5,6. Кроме того адипоциты играют важную роль в регулировании раковых клеток биологии7,8,9,10 и метаболизм4,11,12 ,13,14,,1516, а также в костных метастазов1,4,12. В нише костного мозга адипоциты также может повлиять на биологическое поведение раковых клеток4,6,17. Взаимодействие между адипоциты костного мозга и раковые клетки с osteotropism имеет важное значение для понимания костных метастазов. Однако мало что известно.
Основываясь на текущих исследований, различные методы применяются к адипоциты, включая двух - и трехмерных (2/3D) и ex vivo культур17,18,19,,2021. Herroon et al. , недавно, разработал новый 3D-культура подход к изучению взаимодействия адипоциты костного мозга с раком клетки22. Хотя 3D coculture оптимальное подражая физиологических взаимодействий между адипоциты и рак клеток в естественных условиях, она страдает от бедных воспроизводимость22,23. По сравнению с 2D coculture системы 3D coculture система может обеспечить различных клеточных фенотипы, например клетки морфология21,22,24,25,26. Кроме того ex vivo культуры изолированных губчатой костной ткани фрагментов может привести к надежной нарост адипоцитов искусственный костный мозг клеток17.
В отличие от этих предыдущих моделей однако, 2D ячеечная модель культуры остается классический, надежной и простой метод для быстро сканирования кандидат молекул и фенотипы, изменилось в адипоцитов или рак клеток в пробирке1, 4,6,12,15,27. Чтобы лучше понять уровень помех между адипоциты костного мозга и клетки меланомы, мы предоставляем подробный протокол для 2D coculture системы адипоциты костного мозга с клетки меланомы.

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	<tbody>
		<tr height="17">
			<td height="17" style="height:18px;width:215px;">DMEM</td>
			<td style="width:140px;">Invitrogen Inc.</td>
			<td style="width:143px;">11965092</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Fetal Bovine Serum</td>
			<td style="width:140px;">Invitrogen Inc.</td>
			<td style="width:143px;">16000&#8211;044</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Phosphate Buffered Saline</td>
			<td style="width:140px;">Invitrogen Inc.</td>
			<td style="width:143px;">14190-144</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Insulin</td>
			<td style="width:140px;">Sigma-Aldrich</td>
			<td style="width:143px;">91077C</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">3-isobutyl-1-methyl-xanthine</td>
			<td style="width:140px;">Sigma-Aldrich</td>
			<td style="width:143px;">I5879</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Dexamethasone</td>
			<td style="width:140px;">Sigma-Aldrich</td>
			<td style="width:143px;">D4902</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Oil Red o</td>
			<td style="width:140px;">Sigma-Aldrich</td>
			<td style="width:143px;">O0625</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">24-well plate</td>
			<td style="width:140px;">Corning</td>
			<td style="width:143px;">CLS3527</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Transwell insert</td>
			<td style="width:140px;">Millipore</td>
			<td style="width:143px;">MCHT24H48</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Penicillin/Streptomycin</td>
			<td style="width:140px;">Invitrogen</td>
			<td style="width:143px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">isopropanol</td>
			<td style="width:140px;">Sigma-Aldrich</td>
			<td style="width:143px;">I9516</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">0.25% trypsin</td>
			<td style="width:140px;">Thermo Scientific</td>
			<td style="width:143px;">25200056</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">hemocytometer</td>
			<td style="width:140px;">Bio-Rad</td>
			<td style="width:143px;">1450016</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Culture incubator</td>
			<td style="width:140px;">Thermo Scientific</td>
			<td style="width:143px;"></td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">50ml falcon</td>
			<td style="width:140px;">Corning</td>
			<td style="width:143px;">CLS430828</td>
			<td style="width:187px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Clean Bench</td>
			<td style="width:140px;">Thermo Scientific</td>
			<td style="width:143px;">-</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">Microscopy</td>
			<td style="width:140px;">Olympus</td>
			<td style="width:143px;">-</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">200 &#956;L pipet tips</td>
			<td style="width:140px;">BeyoGold</td>
			<td style="width:143px;">FTIP620</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:215px;">1000 mL pipet tips</td>
			<td style="width:140px;">BeyoGold</td>
			<td style="width:143px;">FTIP628</td>
			<td></td>
		</tr>
	</tbody>
</table>

dual effects of melanoma cell-derived factors on bone marrow adipocytes differentiation

Перекрестных помех между адипоциты костного мозга и клетки опухоли могут сыграть важную роль в процессе костных метастазов. Разнообразные методы доступны для изучения существенных помех; Однако двумерных transwell системы для coculture остается классический, надежный и простой способ для этого исследования помех. Здесь мы представляем подробный протокол, который показывает coculture адипоциты костного мозга и клетки меланомы. Тем не менее такая система coculture может не только способствовать изучение клеток сигнал transductions раковых клеток, вызванные адипоциты костного мозга, но также в будущее механистический исследования костных метастазов, которая может выявить новых терапевтических целей для костей метастазирование.

В костном мозге, адипоциты может появиться в опухоли микроокружения1,13,33,,3435 на ранней стадии для поддержки опухолевой прогрессии через растворимых факторов или Активация osteoclastogen...

Watch this Scientific Journal Video about Dual Effects of Melanoma Cell-derived Factors on Bone Marrow Adipocytes Differentiation at JoVE.com

Dual Effects of Melanoma Cell-derived Factors on Bone Marrow Adipocytes Differentiation

Здесь мы представляем систему надежной и простой двухмерный (2D) coculture для изучения взаимодействия между опухолевых клеток и адипоциты костного мозга, который показывает двойной эффект факторов клеток меланомы на адипоциты костного мозга дифференциация и также представляет классический метод механистический исследования костных метастазов.

Двойное воздействие на дифференциации адипоциты костного факторов клеток меланомы

The crosstalk between bone marrow adipocytes and tumor cells may play a critical role in the process of bone metastasis. A variety of methods are ...

Research

JoVE Journal

Cancer Research

Conditional Knockdown of Gene Expression in Cancer Cell Lines to Study the Recruitment of Monocytes/Macrophages to the Tumor Microenvironment

Условное нокдаун экспрессии генов в рак клеточных линий для изучения набора моноцитов/макрофагов в микроокружения опухоли

siRNA and shRNA-mediated knock down (KD) methods of regulating gene expression are invaluable tools for understanding gene and protein function. However, ...

Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia

Всеобъемлющий протокол выборки и процесс костного мозга для измерения поддающихся оценке остаточной болезни и лейкозных клеток острой миелоидной лейкемией

Response criteria in acute myeloid leukemia (AML) has recently been re-established, with morphologic examination utilized to determine whether patients ...

A 3D Organotypic Melanoma Spheroid Skin Model

Модель 3D Organotypic меланомы кожи сфероида

Malignant transformation of melanocytes, the pigment cells of human skin, causes formation of melanoma, a highly aggressive cancer with increased ...

Multimodal Bioluminescent and Positronic-emission Tomography/Computational Tomography Imaging of Multiple Myeloma Bone Marrow Xenografts in NOG Mice

Смешанных биолюминесцентных и позитронный выбросов томография/вычислительная томография томография множественной миеломы ксенотрасплантатов костного мозга мышей Нагель

Multiple myeloma (MM) tumors engraft in the bone marrow (BM) and their survival and progression are dependent upon complex molecular and cellular ...

A Melanoma Patient-Derived Xenograft Model

Меланома пациента-производные Ксенотрансплантат Модель

Accumulating evidence suggests that molecular and biological properties differ in melanoma cells grown in traditional two-dimensional tissue culture ...

Generation of a Liver Orthotopic Human Uveal Melanoma Xenograft Platform in Immunodeficient Mice

Поколение печени ортотопические человека увеянные меланомы Ксенотрансплантат Платформы в иммунодефицитных мышей

In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more ...

Characterization of the Effects of Migrastatic Inhibitors on 3D Tumor Spheroid Invasion by High-resolution Confocal Microscopy

Характеристика влияния миграстатических ингибиторов на 3D-вторжение опухолевых сфероидов с помощью конфокальной микроскопии высокого разрешения

Drug discovery and development in cancer research is increasingly being based on drug screens in a 3D format. Novel inhibitors targeting the migratory and ...

Modeling Primary Bone Tumors and Bone Metastasis with Solid Tumor Graft Implantation into Bone

Моделирование первичных опухолей костей и метастазов в кости с имплантацией солидного опухолевого трансплантата в кость

Primary bone tumors or bone metastasis from solid tumors result in painful osteolytic, osteoblastic, or mixed osteolytic/osteoblastic lesions. These ...

Mapping the Structure-Function Relationships of Disordered Oncogenic Transcription Factors Using Transcriptomic Analysis

Отображение структурно-функциональных отношений неупорядоченных онкогенных факторов транскрипции с использованием транскриптомного анализа

Many cancers are characterized by chromosomal translocations which result in the expression of oncogenic fusion transcription factors. Typically, these ...

Modeling the Effects of Hemodynamic Stress on Circulating Tumor Cells using a Syringe and Needle

Моделирование влияния гемодинамического стресса на циркулирующие опухолевые клетки с помощью шприца и иглы

During metastasis, cancer cells from solid tissues, including epithelia, gain access to the lymphatic and hematogenous circulation where they are exposed ...