Name: use of a linear accelerator for conducting in vitro radiobiology experiments
Uploaded: 2019-05-26T13:00:00
Description: Watch this Scientific Journal Video about Use of a Linear Accelerator for Conducting In Vitro Radiobiology Experiments at JoVE.com

我们感谢克利夫兰诊所放射肿瘤学系使用线性加速器。我们感谢杰里米·里奇博士慷慨地赠送了胶质瘤干细胞。这项研究得到了克利夫兰诊所的支持。

1. 悬浮细胞培养的细胞制备
<ol><li>在细胞培养箱中培养的干细胞培养基培养基培养基细胞约5×106细胞/10厘米板,CO2为5%,37°C为95%相对湿度。 注: 细胞培养条件在所有过程中都相同。协议中使用的介质是完整的介质。</li>
	<li>在预定辐照前两天,用无菌的5mL移液器从培养板收集胶质瘤干细胞,放入培养罩中的15 mL离心管中。</li>
	<li>在台面离心机中,以 200 x g将收集的细胞离心 3 ?...

<ol>
	<li>Stupp, R., 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=Radiotherapy+plus+concomitant+and+adjuvant+temozolomide+for+glioblastoma.">Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma.</a> The New England Journal of Medicine. 352 (10), 987-996 (2005).</li><li>Stupp, R., 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=Effects+of+radiotherapy+with+concomitant+and+adjuvant+temozolomide+versus+radiotherapy+alone+on+survival+in+glioblastoma+in+a+randomised+phase+III+study:+5-year+analysis+of+the+EORTC-NCIC+trial.">Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial.</a> The Lancet Oncology. 10 (5), 459-466 (2009).</li><li>Tao, R., 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=Hypoxia+imaging+in+upper+gastrointestinal+tumors+and+application+to+radiation+therapy.">Hypoxia imaging in upper gastrointestinal tumors and application to radiation therapy.</a> Journal of Gastrointestinal Oncology. 9 (6), 1044-1053 (2018).</li><li>Gajiwala, S., Torgeson, A., Garrido-Laguna, I., Kinsey, C., Lloyd, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combination+immunotherapy+and+radiation+therapy+strategies+for+pancreatic+cancer-targeting+multiple+steps+in+the+cancer+immunity+cycle.">Combination immunotherapy and radiation therapy strategies for pancreatic cancer-targeting multiple steps in the cancer immunity cycle.</a> Journal of Gastrointestinal Oncology. 9 (6), 1014-1026 (2018).</li><li>Liney, G. P., Whelan, B., Oborn, B., Barton, M., Keall, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI-Linear+accelerator+raiotherapy+systems.">MRI-Linear accelerator raiotherapy systems.</a> Clinical Oncology Journal | The Royal College of Radiologists. 30 (11), 686-691 (2018).</li><li>Hall, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation+dose-rate:+a+factor+of+importance+in+radiobiology+and+radiotherapy.">Radiation dose-rate: a factor of importance in radiobiology and radiotherapy.</a> The British Journal of Radiology. 45 (530), 81-97 (1972).</li><li>Steel, G. G., 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=The+dose-rate+effect+in+human+tumour+cells.">The dose-rate effect in human tumour cells.</a> Radiotherapy &amp; Oncology. 9 (4), 299-310 (1987).</li><li>Ling, C. C., Gerweck, L. E., Zaider, M., Yorke, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose-rate+effects+in+external+beam+radiotherapy+redux.">Dose-rate effects in external beam radiotherapy redux.</a> Radiotherapy &amp; Oncology. 95 (3), 261-268 (2010).</li><li>Castro, G., 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=Amotosalen/UVA+treatment+inactivates+T+cells+more+effectively+than+the+recommended+gammadose+for+prevention+of+transfusion-associated+graft-versus-host+disease.">Amotosalen/UVA treatment inactivates T cells more effectively than the recommended gammadose for prevention of transfusion-associated graft-versus-host disease.</a> Transfusion. 58 (6), 1506-1515 (2018).</li><li>Gaddini, 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=Exposing+primary+rat+retina+cell+cultures+to+&#947;-rays:+An+in+vitro+model+for+evaluating+radiation+responses.">Exposing primary rat retina cell cultures to &#947;-rays: An in vitro model for evaluating radiation responses.</a> Experimental Eye Research. 166, 21-28 (2018).</li><li>Simara, 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=DNA+double-strand+breaks+in+human+induced+pluripotent+stem+cell+reprogramming+and+long-term+in+vitro+culturing.">DNA double-strand breaks in human induced pluripotent stem cell reprogramming and long-term in vitro culturing.</a> Stem Cell Research &amp; Therapy. 8 (1), 73 (2017).</li><li>Wang, 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=A+comparison+of+the+biological+effects+of+125I+seeds+continuous+low-dose-rate+radiation+and+60Co+high-dose-rate+gamma+radiation+on+non-small+cell+lung+cancer+cells.">A comparison of the biological effects of 125I seeds continuous low-dose-rate radiation and 60Co high-dose-rate gamma radiation on non-small cell lung cancer cells.</a> PLoS One. 10 (8), 0133728 (2015).</li><li>Lasio, G., Guerrero, M., Goetz, W., Lima, F., Baulch, 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=Effect+of+varying+dose-per-pulse+and+average+dose+rate+in+X-ray+beam+irradiation+on+cultured+cell+survival.">Effect of varying dose-per-pulse and average dose rate in X-ray beam irradiation on cultured cell survival.</a> Radiation and Environmental Biophysics. 53 (4), 671-676 (2014).</li><li>Karan, 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=Radiobiological+effects+of+altering+dose+rate+in+filter-free+photon+beams.">Radiobiological effects of altering dose rate in filter-free photon beams.</a> Physics in Medicine and Biology. 58 (4), 1075-1082 (2013).</li><li>Sarojini, 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=A+combination+of+high+dose+rate+(10X+FFF/2400+MU/min/10+MV+X-rays)+and+total+low+dose+(0.5+Gy)+induces+a+higher+rate+of+apoptosis+in+melanoma+cells+in+vitro+and+superior+preservation+of+normal+melanocytes.">A combination of high dose rate (10X FFF/2400 MU/min/10 MV X-rays) and total low dose (0.5 Gy) induces a higher rate of apoptosis in melanoma cells in vitro and superior preservation of normal melanocytes.</a> Melanoma Research. 25 (5), 376-389 (2015).</li><li>Hao, 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=The+effects+of+extra+high+on+glioma+stem-like+cells.">The effects of extra high on glioma stem-like cells.</a> PLoS One. 13 (8), 0202533 (2018).</li><li>Liu, 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=Radiation-induced+G2/M+arrest+rarely+occurred+in+glioblastoma+stem-like+cells.">Radiation-induced G2/M arrest rarely occurred in glioblastoma stem-like cells.</a> International Journal of Radiation Biology. 94 (4), 394-402 (2018).</li><li>Mcdermott, 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=.">.</a> The Physics and Technology of Radiation Therapy. , (2010).</li><li>Lohse, I., 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=Effect+of+high+dose+per+pulse+flattening+filter-free+beams+on+cancer+cell+survival.">Effect of high dose per pulse flattening filter-free beams on cancer cell survival.</a> Radiotherapy &amp; Oncology. 101 (1), 226-232 (2011).</li></ol>

放射治疗是癌症管理不可分割的一部分。正在进行的努力旨在提高放射治疗的疗效和效率。直线加速器技术的进步为以前所未有的准确性和安全性治疗患者提供了机会。由于大多数患者都使用直线加速器的高能 X 射线进行治疗,因此,在线性加速器上执行大量剂量率的生物学影响的研究可能很容易应用于患者。已经有一些报告将线性加速器应用于辐射生物学研究,但结果好坏参半,还需要进行13、14、15...

放射治疗是治疗多种癌症的有效手段,治疗1、2、3、4。超高剂量照射在放射治疗中是相对较新的,并且由于线性加速器5的最新技术进步而成为可能。超高剂量率比标准剂量率照射的临床优势包括缩短治疗时间和改善患者体验。线性加速器也为基于细胞培养的辐射生物学研究提供了临床环境。辐射剂量和剂量率的生物和治疗影响一直是放射肿瘤学家和生物学家的兴趣,第6,7,8。但是,超高剂量照射和闪光照射的放射生物学-一个极高的辐射剂量率-还有待彻底调查。
伽马射线照射被广泛应用于基于细胞培养的辐射生物学9、10、11。辐射是通过从衰变的放射性同位素源(通常是Cesium-137)发射的伽马射线来实现的。放射源的使用受到高度管制,而且往往受到限制。使用源辐照,很难测试广泛的剂量率,限制了其在分析临床可实现剂量率12的生物效应的效用。
有几个研究说明剂量和剂量率效应12,13,14,15,16,17。在这些研究中,使用了放射性同位素产生的伽马辐照或线性加速器产生的X射线。使用了代表肺癌、宫颈癌、胶质细胞瘤和黑色素瘤的各种细胞系。辐射对细胞生存、细胞周期抑制、凋亡和DNA损伤的影响被评估为读出12,13,14,15,16,17.在这里,我们描述了一种通过使用线性加速器提供基于 X 射线的辐射来定义临床相关辐射剂量和剂量率的生物效应的方法。这些研究应该由生物学家、放射肿瘤学家和医学物理学家密切合作进行。

<table border="0" cellpadding="0" cellspacing="0" style="width:925px;" width="924">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Material</td>
			<td style="width:269px;"></td>
			<td style="width:183px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">glioma stem-like cell 4121</td>
			<td style="width:269px;"></td>
			<td style="width:183px;"></td>
			<td>gift from Dr. Jeremy Rich</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">293 cells</td>
			<td style="width:269px;">ATCC</td>
			<td style="width:183px;">CRL-1573</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:307px;">neuron stem cell culture media</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">21103049</td>
			<td>NeurobasalTM media</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">DMEM</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">10569044</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Fetal Bovine Serum</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">16000044</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Penicillin/Streptomycin</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">15140-122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Recombinant Human EGF Protein</td>
			<td style="width:269px;">R&#38;D Systems</td>
			<td>236-EG-01M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Recombinant Human FGF basic</td>
			<td style="width:269px;">R&#38;D Systems</td>
			<td>4114-TC-01M</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">B-27&#8482; Supplement</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">17504044</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Pyruvate</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">11360070</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">L-Glutamine</td>
			<td style="width:269px;">Thermo Fisher Scientific</td>
			<td style="width:183px;">25030164</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Tripsin-EDTA</td>
			<td style="width:269px;">Thermo Fisher</td>
			<td style="width:183px;">25200056</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:307px;">extracellular proten matrix</td>
			<td style="width:269px;">Corning</td>
			<td style="width:183px;">354277</td>
			<td style="width:167px;">MatrigelTM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Ethanol</td>
			<td style="width:269px;">Fisher chemical</td>
			<td style="width:183px;">A4094</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Equipment</td>
			<td style="width:269px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">10 cm cell culture dish</td>
			<td style="width:269px;">Denville</td>
			<td style="width:183px;">T1110</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">3.5 cm cell culture dish</td>
			<td style="width:269px;">USA Scientific Inc.</td>
			<td style="width:183px;">CC7682-3340</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">22x22mm glass cover slip</td>
			<td style="width:269px;">electron microscopy sciences</td>
			<td style="width:183px;">72210-10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">15 ml centrifuge tube</td>
			<td style="width:269px;">Thomas Scientific</td>
			<td style="width:183px;">1159M36</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">50 ml centrifuge tube</td>
			<td style="width:269px;">Thomas Scientific</td>
			<td style="width:183px;">1158R10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">5 ml Pipette</td>
			<td style="width:269px;">Fisher Scientific</td>
			<td style="width:183px;">14-955-233</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">pipet aid</td>
			<td style="width:269px;">Fisher Scientific</td>
			<td style="width:183px;">13-681-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Vortex mixer</td>
			<td style="width:269px;">Fisher Scientific</td>
			<td style="width:183px;">02-215-414</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Centrifuge</td>
			<td style="width:269px;">Eppendorf</td>
			<td style="width:183px;">5810R</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Linear Accelerator</td>
			<td style="width:269px;">Varian</td>
			<td style="width:183px;">n/a</td>
			<td></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:307px;">water equivalent material</td>
			<td style="width:269px;">Sun Nuclear corporation</td>
			<td style="width:183px;">557</td>
			<td>Solid waterTM</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Reagent preparation</td>
			<td style="width:269px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:307px;">DMEM media</td>
			<td style="width:269px;">10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/mL penicillin G, 100 &#181;g/mL streptomycin in 500 ml DMEM media</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="105">
			<td height="105" style="height:105px;width:307px;">stem cell culture media</td>
			<td style="width:269px;">10 ml B27 supplement, 20 &#181;g hFGF, 20 &#181;g hEGF, 2 mM L-glutamine, 100 units/mL penicillin G, 100 &#181;g/mL streptomycin in 500 ml Neurobasal media</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

use of a linear accelerator for conducting in vitro radiobiology experiments

放射治疗仍然是癌症管理的基石之一。对于大多数癌症,它是最有效的,非手术治疗去散体肿瘤。在这里,我们描述了一种用线性加速器照射癌细胞的方法。直线加速器技术的发展提高了放射治疗的精度和效率。广泛的辐射剂量和剂量率的生物影响仍然是一个紧张的研究领域。使用线性加速器可以促进这些研究使用临床相关的剂量和剂量率。

为了研究标准剂量率和线性加速器照射超高剂量率的细胞周期效应,使用该协议制备了三个胶质瘤干细胞样本,并在辐照17后收集了24小时:一个对照样本未辐照(图2A),一个样品照射400 MU/min(监测单位,4.2 Gy/min标准剂量率,图2B)至4 Gy,另一个样品照射2100 MU/min(21.2 Gy/min超高剂量率,图 2C...

Watch this Scientific Journal Video about Use of a Linear Accelerator for Conducting In Vitro Radiobiology Experiments at JoVE.com

Use of a Linear Accelerator for Conducting In Vitro Radiobiology Experiments

使用线性加速器进行体外放射生物学实验

临床直线加速器可用于确定各种剂量率对癌细胞的生物效应。我们讨论如何为癌症干细胞样细胞的测定和测定建立线性加速器,这些细胞在悬浮物中生长为肿瘤球体,细胞系作为附体培养体生长。

Radiation therapy remains one of the cornerstones of cancer management. For most cancers, it is the most effective, nonsurgical therapy to debulk tumors. ...

放射治疗最常使用线性加速器进行。该协议允许研究人员使用线性加速器评估辐射对细胞的生物影响。该技术的优点是，通过使用线性加速器，研究人员可以研究广泛的临床相关剂量和剂量率。此外，该协议可用于所有类型的癌细胞。此方法可以使用与此处所示类似的设备执行。例如，由其他供应商制造的线性加速器。没有使用线性加速器进行剂量计算经验的个人可能难以计算向样品提供所需剂量所需的正确监测单位数。他们将受益于寻求经验医学物理学家或医生的建议。使用线性加速器准确向样品提供剂量需要仔细放置。在预定辐照前两天，在培养罩中工作，使用无菌的五毫升移液器将胶质瘤干细胞从培养板收集到15毫升离心管中。在台式离心机中以200倍g离心，三分钟。丢弃上一提液，用一毫升的三辛-EDTA重新暂停细胞颗粒。在室温下孵育五分钟，消化颗粒，并在尝试性-EDTA中进行单细胞悬浮。在消化过程中，每两分钟轻轻摇动离心管底部，以确保细胞被彻底消化。然后加入三毫升干细胞培养培养剂，在台面离心机中以200倍g为单位轻轻混合并离心机3分钟。丢弃上清液后，用五毫升细胞培养培养剂重新暂停细胞颗粒，用血细胞计对细胞进行计数。板500万细胞分成两个10厘米的板，含有10毫升的细胞培养基，在37摄氏度下孵育48小时。就在预定的辐照之前，收集细胞和离心机，如前完成。丢弃上一提液，用五毫升细胞培养培养剂重新暂停细胞颗粒。将一毫升细胞悬浮液转移到一个35毫米的板中，其中包含两毫升的细胞培养基，确保培养基达到一厘米的高度。将镀有细胞转移到辅助容器中，以降低污染风险，并将公用车上容器中的细胞带到辐照设施，以照射细胞。在照射前一天，在细胞培养罩中工作，使用连接到真空的巴斯德移液器将DMEM培养基从细胞培养板中去除至所附细胞系。然后用五毫升无菌PBS清洗细胞，冲洗掉残留介质。在培养皿中加入一毫升的三辛-EDTA，轻轻倾斜培养盘，确保覆盖整个盘子。在室温下孵育五分钟后，用三毫升DMEM介质淬火抑制三辛-EDTA反应。使用五毫升移液器将细胞收集到15毫升离心管、离心机中，然后培养手稿中描述的细胞。使用 LINAC 的控制台软件照射这些以前准备的细胞，将加速器龙门和准直器设置为零度，将 x 和 y 钳口打开到对称的五乘五平方厘米的场大小，并收回多叶准直直器。在治疗沙发上添加至少五厘米的水当量材料，并将细胞盘照射到 LINAC 十字线的中心。将细胞以最大剂量的深度放在约1.5厘米的6兆伏X射线束中，并在培养皿顶部添加1厘米的水当量材料。将前指针固定到 LINAC 的头部并扩展它，直到它接触堆积材料的表面。调整表高度，直到从源到堆积表面的距离为 100 厘米。离开治疗库，确保所有其他人员已经离开。验证房间中是否有其他单元格，然后关闭保管库门。在控制台上确认场大小、监控单元和每分钟监控单位，然后为辐射单元启用光束。使用该协议编写了三个胶质瘤干细胞样本，并在照射后24小时采集。计算细胞周期配置文件，并显示细胞在细胞周期的不同阶段的百分比。与非辐照控制细胞相比，在以标准剂量率或超高剂量率照射胶质瘤干细胞后观察到G2细胞周期逮捕。为了提供正确的剂量，最重要的是测量培养皿中介质的高度，以确保其达到一厘米。按照这个程序，可以进行其他生物测定，如免疫污染，西部印迹和细胞周期分析。由于该程序为研究人员提供了临床相关的剂量和剂量率设置，它可用于评估辐射效应对许多细胞培养为基础的模型。

Research

JoVE Journal

Medicine

Use of a Hanging Weight System for Coronary Artery Occlusion in Mice

悬重系统的使用在小鼠冠状动脉闭塞

Murine studies of acute injury are an area of intense investigation, as knockout mice for different genes are becoming increasingly available 1-38. ...

科研

医学

Use of a Hanging-weight System for Isolated Renal Artery Occlusion

一个悬重系统使用隔离肾动脉闭塞

In hospitalized patients, over 50% of cases of acute kidney injury (AKI) are caused by renal ischemia 1-3. A recent study of hospitalized patients ...

Use of a Hanging-weight System for Liver Ischemia in Mice

使用吊重系统在小鼠肝缺血

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and ...

The Use of Cystometry in Small Rodents: A Study of Bladder Chemosensation

的的膀胱测压小老鼠：A膀胱Chemosensation研究的使用

	The lower urinary tract (LUT) functions as a  dynamic reservoir that is able to store urine and to efficiently expel it at a  convenient time. While ...

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression

异型三维<em在体外</em&gt;卵巢癌的发生和发展过程中的间质 - 上皮细胞相互作用模型

Epithelial ovarian cancers (EOCs) are the leading cause of death from gynecological malignancy in Western societies. Despite advances in surgical ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Setting Up a Stroke Team Algorithm and Conducting Simulation-based Training in the Emergency Department - A Practical Guide

建立中风病研究小组算法并进行急诊科基于仿真的培训 - 实用指南

Time is of the essence when caring for an acute stroke patient. The ultimate goal is to restore blood flow to the ischemic brain. This can ...

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

Preparation of Herbal Medicine: Er-Xian Decoction and Er-Xian-containing Serum for In Vivo and In Vitro Experiments

草药的制备：洱西汤和含茜素的血清<em&gt;体内</em&gt;和<em&gt;体外</em&gt;实验

Traditional herbal medicine, an alternative medicine in the clinical setting, has received increased attention in recent years. Before delivery to the ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

进行最大和次极大耐力运动测试, 测量人类急性运动的生理和生物学反应

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Preparation Of Gushukang (GSK) Granules for In Vivo and In Vitro Experiments

Preparation Of Gushukang GSK Granules for In Vivo and In Vitro Experiments

古树康 (GSK) 颗粒的制备,用于 Vivo 和体外实验

Traditional Chinese herbal medicine plays a role as an alternative method in treating many diseases, such as postmenopausal osteoporosis (POP). Gushukang ...