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

Biz Dov Zipori (Weizmann Enstitüsü bilim, Rehovot, İsrail) lütfen bize fare kemik iliği stromal hücre verdiğiniz için teşekkür ederiz 14F1.1 hattı. Bu çalışmada hibe Çin Ulusal Doğal Bilim Vakfı (NOS 81771729) ve Yongchuan hastane Chongqing Tıp Üniversitesi ('ları tarafından desteklenmiştir YJQN201330; YJZQN201527).

Not: Bu protokol için kullanılan tüm hücre donmuş hisse senedi hücrelerden çözdürme sonra en az üç nesiller için yetişkin olmalıdır.
1. hasat Melanoma hücre kaynaklı faktörler
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Hazırlıklar
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<li>B16F10 hücreleri ile bir fare melanoma hücre kültürünü edinmek. Not: Bu iletişim kuralı için bir fare melanoma hücre kültürünü Çin Bilimler Akademisi kök hücre bankadan elde edildi.</li>
		<li>B16F10 hücre kültürü (100 mL) için...

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Cocultures ekler ile hücre hücre etkileşimleri eğitim için yaygın olarak kullanılmaktadır. 2D coculture sistemi nasıl iki parça crosstalk vitro, biz burada iki farklı kanser hücre tahrik etkileri kemik iliği adipositler tarafından ortaya gözlemlemek için etkili bir yoldur. Birçok labs adiposit ve kanser hücreleri6,12,27,39arasında çapraz karışma araştırmak için...

Kemik metastazı ileri kanser hastaları arasında yaygın, ancak bir iyileştirici tedavi hala ulaşılamıyor. Yağ olarak enerji depolama uzmanlaşmış ötesinde, adipositler tümör büyüme ve metastaz kemik iliği ve makarna diğer organları1,2,3,4,5,6' da destekleyebilir. Ayrıca, adipositler kanser hücre biyolojisi7,8,9,10 ve metabolizma4,11,12 düzenlenmesinde önemli bir rol oynamak su kuyusu içinde olarak kemik metastaz1,4,12gibi ,13,14,15,16,. Kemik iliği niş içinde adiposit da kanser hücreleri4,6,17biyolojik davranışını etkileyebilir. Kemik iliği adiposit ve osteotropism kanseri hücreleri arasındaki etkileşimi bir kemik metastaz anlamak için önemlidir. Ancak, az şey bilinmektedir.
Mevcut çalışmalar üzerinde bağlı olarak, iki veya üç boyutlu (2/3D) ve ex vivo kültürler17,18,19,20,21de dahil olmak üzere adiposit için çeşitli yöntemler uygulanır. Son zamanlarda, Herroon vd. kemik iliği adiposit etkileşimleri kanser hücreleri22ile çalışmaya yeni bir 3D-kültür yaklaşım tasarlanmış. 3D coculture fizyolojik taklit için en uygun olmasına rağmen adiposit ve kanser arasındaki etkileşimler hücreleri vivo içindebu zavallı tekrarlanabilirlik22,23uğrar. 2D coculture sistemi ile karşılaştırıldığında, bir 3D coculture sistemi gibi hücre morfolojisi21,22,24,25,26farklı hücresel fenotipleri sağlayabilir. Ayrıca, izole Süngerimsi Kemik doku parçaları ex vivo kültürünü adiposit sağlam bir akıbet kültürlü kemik iliği hücreleri17yol açabilir.
Hızlı bir şekilde tarama aday molekülleri ve adiposit veya kanser değişti fenotipleri vitro1hücreleri bu önceki modellerin aksine Ancak, 2D hücre kültür modeli bir klasik, güvenilir ve kolay teknik kalır, 4,6,12,15,27. Kemik iliği adiposit ve Melanom hücreleri arasında çapraz karışma daha iyi anlamak için biz bir detaylı protokol kemik iliği adiposit Melanom hücreleri ile 2D coculture sistemi için sağlar.

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			<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>
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			<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>
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			<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>
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		<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

Kemik iliği adiposit ve tümör hücreleri arasında çapraz karışma kemik metastaz sürecinde önemli bir rol oynayabilir. Çeşitli yöntemler önemli crosstalk eğitim için kullanılabilir; Ancak, bir iki boyutlu transwell sistemi coculture için bir klasik, güvenilir ve kolay bir şekilde bu çapraz karışma çalışma için kalır. Burada, coculture kemik iliği adiposit ve Melanom hücreleri gösterir ayrıntılı bir iletişim kuralı mevcut. Yine de, böyle bir coculture sistem sadece hücre sinyal transductions kemik iliği adipositler tarafından indüklenen kanser hücrelerinin incelenmesi katkıda ama Ayrıca kemik için yeni tedavi hedefleri ortaya çıkarabilir kemik metastaz, geleceğe mekanik çalışma metastaz.

Kemik iliğinde adiposit tümör microenvironment1,13,33,34,35 erken bir aşamada tümör ilerleme çözünür faktörler ile desteklemek için görünebilir veya osteoclastogenesis6,12,36, özellikle obezite6<s...

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

Burada, bir çift etkisi melanoma hücre kaynaklı faktörler kemik iliği adiposit ortaya koymaktadır tümör hücreleri ve kemik iliği adipositler, arasındaki etkileşimi çalışmak için güvenilir ve doğru sözlü iki boyutlu (2D) coculture sistemi mevcut farklılaşma ve ayrıca kemik metastaz mekanik incelenmesi için klasik bir yöntem teşkil etmektedir.

Melanoma hücre kaynaklı faktörler çift etkileri kemik iliği adiposit farklılaşması üzerine

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

Monosit/makrofajlar tümör Microenvironment için işe alım çalışmaya kanser hücre hatlarında gen ifadesinin koşullu nakavt

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

Örnek ve işlem kemik iliği ölçülebilir artık hastalık ve akut Myeloid lösemi lösemik kök hücre ölçmek için kapsamlı iletişim kuralı

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

3D Organotypic Melanom küresel Cilt modeli

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

Multipl Miyelom kemik iliği Xenografts NOG farelerde Multimodal kollarındaki ve pozitronik emisyon tomografi/Hesaplamalı tomografi görüntüleme

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

Bir melanom hasta-türetilen xenograft model

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

İmmünficient Farelerde Karaciğer Ortotopik İnsan Uveal Melanom Xenograft Platformu Üretimi

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

Migrastatik İnhibitörlerin 3Boyutlu Tümör Spheroid İnvazyonu Üzerine Etkilerinin Yüksek Çözünürlüklü Konfokal Mikroskopi Ile Karakterizasyonu

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

Primer Kemik Tümörleri ve Kemik Metastazlarının Solid Tümör Grefti İmplantasyonu ile Modellenmesi

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

Transcriptomik Analiz Kullanarak Düzensiz Onkojenik Transkripsiyon Faktörlerinin Yapı-fonksiyon İlişkilerinin Eşlenmesini

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

Hemodinamik Stresin Dolaşımdaki Tümör Hücreleri Üzerindeki Etkilerini şırınga ve İğne Kullanarak Modelleme

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