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

Ringraziamo gentilmente Dov Zipori (The Weizmann Institute of Science, Rehovot, Israele) per averci fornito le cellule stromali del midollo osseo murino linea 14F1.1. Questo studio è stato sostenuto da sovvenzioni da Fondazione Scienza naturale nazionale cinese (nn. 81771729) e l'Università ospedale di Chongqing Yongchuan medica (Nos. YJQN201330; YJZQN201527).

Nota: Tutte le celle utilizzate nel presente protocollo dovrebbero essere cresciute da almeno tre generazioni dopo lo scongelamento da cellule stock congelate.
1. raccogliere i fattori derivati da cellule di Melanoma
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Preparazioni
	<ol>
<li>Ottenere le cellule B16F10 e una linea cellulare di melanoma del topo. Nota: Per questo protocollo, una linea cellulare di melanoma del topo è stata ottenuta dalla banca della cellula formativa dell'Accademia cinese delle scie...

<ol>
	<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. 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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. 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Cocultures con inserti sono stati ampiamente usati per studiare le interazioni cellula--cellula. Il sistema di coculture 2D è un modo efficace per osservare come le due parti diafonia in vitro, mediante il quale abbiamo mostrato qui due differenti del cancro delle cellule-guidati effetti sugli adipociti del midollo osseo. Molti laboratori hanno utilizzato questo metodo per indagare il crosstalk tra adipociti e cancro cellule6,12,<sup class="x...

Le metastasi dell'osso sono diffuse fra i pazienti di cancro avanzato, ma un trattamento curativo non è ancora disponibile. Di là di specializzata nella conservazione di energia come grasso, adipociti è in grado di supportare la crescita del tumore e metastasi nel midollo osseo e altri organi1,2,3,4,5,6. Inoltre, adipociti svolgono un ruolo essenziale nella regolazione del cancro cellula biologia7,8,9,10 e metabolismo4,11,12 ,13,14,15,16, come bene come in osso metastasi1,4,12. Nella nicchia del midollo osseo, adipociti possono anche influenzare il comportamento biologico del cancro cellule4,6,17. L'interazione tra adipociti del midollo osseo e le cellule tumorali con osteotropism è significativo per la comprensione della metastasi dell'osso. Tuttavia, piccolo è conosciuto.
Basati sugli studi di correnti, vari metodi vengono applicati ai adipocytes, tra cui bi - o tri - dimensionale (2/3D) ed ex vivo culture17,18,19,20,21. Recentemente, Herroon et al. progettato un nuovo approccio di 3D-cultura per studiare le interazioni degli adipociti del midollo osseo con cancro cellule22. Anche se il 3D coculture è ottimale per mimare fisiologiche interazioni tra adipociti e cancro cellule in vivo, soffre di scarsa riproducibilità22,23. Rispetto ad un sistema di coculture 2D, un sistema di coculture 3D può fornire diversi fenotipi cellulari, come ad esempio cellule morfologia21,22,24,25,26. Inoltre, la cultura ex vivo di frammenti di tessuto isolato cancellous dell'osso può portare a un'escrescenza robusta degli adipociti dal midollo osseo coltivate cellule17.
In contrasto con questi modelli precedenti, tuttavia, il modello della coltura cellulare 2D rimane una tecnica classica, affidabile e facile scansione rapidamente molecole candidate e i fenotipi cambiati in adipociti o cancro cellule in vitro1, 4,6,12,15,27. Per comprendere meglio il crosstalk tra adipociti del midollo osseo e le cellule del melanoma, forniamo un protocollo dettagliato per un sistema di coculture 2D degli adipociti del midollo osseo con cellule di melanoma.

<table border="0" cellpadding="0" cellspacing="0" style="width:684px;" width="685">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<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

Il crosstalk tra adipociti del midollo osseo e le cellule del tumore può giocare un ruolo critico nel processo di metastasi ossee. Una varietà di metodi sono disponibili per studiare il crosstalk significativo; Tuttavia, un sistema di transwell bidimensionale per coculture rimane un classico, affidabile e facile modo per questo studio di diafonia. Qui, presentiamo un protocollo dettagliato che mostra il coculture delle adipociti del midollo osseo e le cellule del melanoma. Tuttavia, un tale sistema di coculture potrebbe non solo contribuire allo studio di trasduzioni segnale cellulare delle cellule di cancro indotte dagli adipociti del midollo osseo, ma anche per il futuro meccanicistico studio della metastasi dell'osso che può rivelare nuovi bersagli terapeutici per osso metastasi.

Nel midollo osseo, adipociti possono apparire nel tumore microambiente1,13,33,34,35 in una fase precoce per sostenere la progressione del tumore attraverso fattori solubili o attivazione12,6,osteoclastogenesis36, soprattutto nel contes...

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

Qui, presentiamo un sistema affidabile e semplice coculture bidimensionale (2D) per studiare l'interazione tra le cellule del tumore e adipociti del midollo osseo, che rivela un duplice effetto di fattori derivati da cellule di melanoma su adipociti del midollo osseo differenziazione e pone anche un metodo classico per lo studio meccanicistico della metastasi dell'osso.

Effetti doppi di fattori derivati da cellule di Melanoma su differenziazione adipociti del midollo osseo

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

Atterramento condizionale dell'espressione genica in linee cellulari del cancro per studiare il reclutamento dei monociti/macrofagi al microambiente tumorale

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

Ricerca

Ricerca sul cancro

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

Protocollo completo del campione e del midollo osseo di processo per misurare la malattia residua misurabile e cellule staminali leucemiche nella leucemia mieloide acuta

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

Un modello di pelle sferoide 3D Organotypic Melanoma

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

Formazione immagine di tomografia multimodale bioluminescenti ed emissione positronica tomografia/computazionale di mieloma multiplo del midollo osseo xenotrapianti in topi NOG

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

Un modello di Xenotrapianto a parte del paziente di melanoma

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

Generazione di una piattaforma di Melanoma Uvenoma Uvenoft umano del fegato in topi immunodeficienti

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

Caratterizzazione degli effetti degli inibitori espastatici sull'invasione 3D dello spheroid del tumore mediante microscopia confocale ad alta risoluzione

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

Modellazione dei tumori ossei primari e delle metastasi ossee con impianto di innesto tumorale solido nell'osso

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

Mappatura delle relazioni struttura-funzione dei fattori di trascrizione oncogenica disordinati utilizzando l'analisi trascrittomica

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

Modellazione degli effetti dello stress emodinamico sulle cellule tumorali circolanti utilizzando una siringa e un ago

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