<meta charset="utf8"></head><body>使用循环肿瘤细胞研究软脑膜疾病的小鼠模型

<ol>
	<li>Glitza, I. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leptomeningeal+disease+in+melanoma+patients:+An+update+to+treatment,+challenges,+and+future+directions.">Leptomeningeal disease in melanoma patients: An update to treatment, challenges, and future directions.</a> Pigment Cell Melanoma Res. 33 (4), 527-541 (2020).</li><li>Khaled, M. L., Tarhini, A. A., Forsyth, P. A., Smalley, I., Pina, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leptomeningeal+disease+(LMD)+in+patients+with+melanoma+metastases.">Leptomeningeal disease (LMD) in patients with melanoma metastases.</a> Cancers (Basel). 15 (6), 1884 (2023).</li><li>Smalley, 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=Proteomic+analysis+of+CSF+from+patients+with+leptomeningeal+melanoma+metastases+identifies+signatures+associated+with+disease+progression+and+therapeutic+resistance).">Proteomic analysis of CSF from patients with leptomeningeal melanoma metastases identifies signatures associated with disease progression and therapeutic resistance).</a> Clin Cancer Res. 26 (9), 2163-2175 (2020).</li><li>Larkin, 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=Five-year+survival+with+combined+nivolumab+and+ipilimumab+in+advanced+melanoma.">Five-year survival with combined nivolumab and ipilimumab in advanced melanoma.</a> N Engl J Med. 381, 1535-1546 (2019).</li><li>Boire, 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=Complement+component+3+adapts+the+cerebrospinal+fluid+for+leptomeningeal+metastasis.">Complement component 3 adapts the cerebrospinal fluid for leptomeningeal metastasis.</a> Cell. 168 (6), 1101-1113 (2017).</li><li>Chi, 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=Cancer+cells+deploy+lipocalin-2+to+collect+limiting+iron+in+leptomeningeal+metastasis.">Cancer cells deploy lipocalin-2 to collect limiting iron in leptomeningeal metastasis.</a> Science. 369 (6501), 276-282 (2020).</li><li>Law, V., 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+preclinical+model+of+patient-derived+cerebrospinal+fluid+circulating+tumor+cells+for+experimental+therapeutics+in+leptomeningeal+disease+from+melanoma.">A preclinical model of patient-derived cerebrospinal fluid circulating tumor cells for experimental therapeutics in leptomeningeal disease from melanoma.</a> Neuro Oncol. 24 (10), 1673-1686 (2022).</li><li>Carmona-Ule, 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=Short-term+ex+vivo+culture+of+CTCs+from+advance+breast+cancer+patients:+Clinical+implications.">Short-term ex vivo culture of CTCs from advance breast cancer patients: Clinical implications.</a> Cancers (Basel). 13 (11), 2668 (2021).</li><li>Zhang, 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=The+identification+and+characterization+of+breast+cancer+CTCs+competent+for+brain+metastasis.">The identification and characterization of breast cancer CTCs competent for brain metastasis.</a> Sci Transl Med. 5 (180), (2013).</li><li>Yu, 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=Cancer+therapy:+Ex+vivo+culture+of+circulating+breast+tumor+cells+for+individualized+testing+of+drug+susceptibility.">Cancer therapy: Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility.</a> Science. 345 (6193), 216-220 (2014).</li><li>Mohamed, B. 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=Ex+vivo+expansion+of+circulating+tumour+cells+(CTCs).">Ex vivo expansion of circulating tumour cells (CTCs).</a> Sci Rep. 13 (1), 3704 (2023).</li><li>Decimo, I., Fumagalli, G., Berton, V., Krampera, M., Bifari, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Meninges:+From+protective+membrane+to+stem+cell+niche.">Meninges: From protective membrane to stem cell niche.</a> Am J Stem Cells. 1 (2), 92-105 (2012).</li><li>Mercier, F., Hatton, G. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Connexin+26+and+basic+fibroblast+growth+factor+are+expressed+primarily+in+the+subpial+and+subependymal+layers+in+adult+brain+parenchyma:+Roles+in+stem+cell+proliferation+and+morphological+plasticity.">Connexin 26 and basic fibroblast growth factor are expressed primarily in the subpial and subependymal layers in adult brain parenchyma: Roles in stem cell proliferation and morphological plasticity.</a> J Comp Neurol. 431 (1), 88-104 (2001).</li><li>Stylianopoulou, F., Herbert, J., Soares, M. B., Efstratiadis, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+the+insulin-like+growth+factor+II+gene+in+the+choroid+plexus+and+the+leptomeninges+of+the+adult+rat+central+nervous+system.">Expression of the insulin-like growth factor II gene in the choroid plexus and the leptomeninges of the adult rat central nervous system.</a> Proc Natl Acad Sci U S A. 85 (1), 141-145 (1988).</li><li>Nordqvist, A. C., Mathiesen, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+IGF-II,+IGFBP-2,+-5,+and+-6+in+meningiomas+with+different+brain+invasiveness.">Expression of IGF-II, IGFBP-2, -5, and -6 in meningiomas with different brain invasiveness.</a> J Neurooncol. 5, 19-26 (2002).</li><li>Zumkeller, W., Westphal, 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+IGF/IGFBP+system+in+CNS+malignancy.">The IGF/IGFBP system in CNS malignancy.</a> Mol Pathol. 54 (4), 227-229 (2001).</li><li>Wang, 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=Promise+and+limits+of+the+CellSearch+platform+for+evaluating+pharmacodynamics+in+circulating+tumor+cells.">Promise and limits of the CellSearch platform for evaluating pharmacodynamics in circulating tumor cells.</a> Semin Oncol. 43 (4), 464-475 (2016).</li><li>Liu, 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=Patient-derived+xenograft+models+in+cancer+therapy:+technologies+and+applications.">Patient-derived xenograft models in cancer therapy: technologies and applications.</a> Signal Transduct Target Ther. 8, 160 (2023).</li><li>Chen, 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+development+and+improvement+of+immunodeficient+mice+and+humanized+immune+system+mouse+models.">The development and improvement of immunodeficient mice and humanized immune system mouse models.</a> Front Immunol. 13, 1007579 (2022).</li><li>Law, V., 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+Murine+Ommaya+xenograft+model+to+study+direct-targeted+therapy+of+leptomeningeal+disease.">A Murine Ommaya xenograft model to study direct-targeted therapy of leptomeningeal disease.</a> J Vis Exp. (167), e62033 (2021).</li><li>Stacer, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NanoLuc+reporter+for+dual+luciferase+imaging+in+living+animals.">NanoLuc reporter for dual luciferase imaging in living animals.</a> Mol Imaging. 12 (7), 1-13 (2013).</li><li>Luo, Y. 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=The+viable+circulating+tumor+cells+with+cancer+stem+cells+feature,+where+is+the+way+out.">The viable circulating tumor cells with cancer stem cells feature, where is the way out.</a> J Exp Clin Cancer Res. 37, 38 (2018).</li><li>Stumm, R., Kolodziej, A., Schulz, S., Kohtz, J. D., Hollt, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterns+of+SDF-1alpha+and+SDF-1gamma+mRNAs,+migration+pathways,+and+phenotypes+of+CXCR4-expressing+neurons+in+the+developing+rat+telencephalon.">Patterns of SDF-1alpha and SDF-1gamma mRNAs, migration pathways, and phenotypes of CXCR4-expressing neurons in the developing rat telencephalon.</a> J Comp Neurol. 502 (3), 382-399 (2007).</li><li>Aviezer, 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=basal+lamina+proteoglycan,+promotes+basic+fibroblast+growth+factor-receptor+binding,+mitogenesis,+and+angiogenesis.">basal lamina proteoglycan, promotes basic fibroblast growth factor-receptor binding, mitogenesis, and angiogenesis.</a> Cell. 79 (6), 1005-1013 (1994).</li><li>Tiwary, 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=ERBB3+is+required+for+metastasis+formation+of+melanoma+cells.">ERBB3 is required for metastasis formation of melanoma cells.</a> Oncogenesis. 3 (7), e110 (2014).</li><li>Sun, X., 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=miR-7+reverses+the+resistance+to+BRAFi+in+melanoma+by+targeting+EGFR/IGF-1R/CRAF+and+inhibiting+the+MAPK+and+PI3K/AKT+signaling+pathways.">miR-7 reverses the resistance to BRAFi in melanoma by targeting EGFR/IGF-1R/CRAF and inhibiting the MAPK and PI3K/AKT signaling pathways.</a> Oncotarget. 7 (33), 53558-53570 (2016).</li><li>Satyamoorthy, K., Li, G., Vaidya, B., Patel, D., Herlyn, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin-like+growth+factor-1+induces+survival+and+growth+of+biologically+early+melanoma+cells+through+both+the+mitogen-activated+protein+kinase+and+beta-catenin+pathways.">Insulin-like growth factor-1 induces survival and growth of biologically early melanoma cells through both the mitogen-activated protein kinase and beta-catenin pathways.</a> Cancer Res. 61 (19), 7318-7324 (2001).</li><li>Lin, N. U., 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=Tucatinib+vs+Placebo,+both+in+combination+with+Trastuzumab+and+Capecitabine,+for+previously+treated+ERBB2+(HER2)-positive+metastatic+breast+cancer+in+patients+with+brain+metastases:+Updated+exploratory+analysis+of+the+HER2CLIMB+randomized+clinical+trial.">Tucatinib vs Placebo, both in combination with Trastuzumab and Capecitabine, for previously treated ERBB2 (HER2)-positive metastatic breast cancer in patients with brain metastases: Updated exploratory analysis of the HER2CLIMB randomized clinical trial.</a> JAMA Oncol. 9 (2), 197-205 (2023).</li><li>Russo, 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=Ceritinib-induced+regression+of+an+insulin-like+growth+factor-driven+neuroepithelial+brain+tumor.">Ceritinib-induced regression of an insulin-like growth factor-driven neuroepithelial brain tumor.</a> Int J Mol Sci. 20 (17), 4267 (2019).</li><li>Ohsie, S. J., Sarantopoulos, G. P., Cochran, A. J., Binder, S. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immunohistochemical+characteristics+of+melanoma.">Immunohistochemical characteristics of melanoma.</a> J Cutan Pathol. 35 (5), 433-444 (2008).</li><li>Kulasinghe, 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=Short+term+ex-vivo+expansion+of+circulating+head+and+neck+tumour+cells.">Short term ex-vivo expansion of circulating head and neck tumour cells.</a> Oncotarget. 7 (37), 60101-60109 (2016).</li><li>Li, X., 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=Clinical+significance+of+detecting+CSF-derived+tumor+cells+in+breast+cancer+patients+with+leptomeningeal+metastasis.">Clinical significance of detecting CSF-derived tumor cells in breast cancer patients with leptomeningeal metastasis.</a> Oncotarget. 9 (2), 2705-2714 (2018).</li><li>Lelliott, E. 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=A+novel+immunogenic+mouse+model+of+melanoma+for+the+preclinical+assessment+of+combination+targeted+and+immune-based+therapy.">A novel immunogenic mouse model of melanoma for the preclinical assessment of combination targeted and immune-based therapy.</a> Sci Rep. 9 (1), 1225 (2019).</li></ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL syringe 27 - 29 G needles</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td>0.1 mm Sterile Filtered</td>
		</tr>
		<tr>
			<td>1.5 mL Eppendorf tubes</td>
			<td>Any vendor</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>15 ml and 50 mL polystyrene centrifuge tubes</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>6 -&#160; 8 weeks females NOD SCID gamma (NSG) mice</td>
			<td>Charles River</td>
			<td></td>
			<td>Males optional</td>
		</tr>
		<tr>
			<td>Buprenorphine Sustained-Release (Bup-SR)</td>
			<td>Zoopharm&#160;</td>
			<td></td>
			<td>DEA controlled</td>
		</tr>
		<tr>
			<td>Fetal bovine serum (FBS)</td>
			<td>ScienCell</td>
			<td>#0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Gas inhalation anestehsia system</td>
			<td>VeteEquip</td>
			<td>#901812</td>
			<td>COMPAC5</td>
		</tr>
		<tr>
			<td>Hamilton microliter syringes</td>
			<td>Hamilton</td>
			<td>10, 25, 50, and 100ml</td>
			<td>30 G for cisterna magna injection</td>
		</tr>
		<tr>
			<td>Human basic fibroblast growth factor (bFGF)</td>
			<td>Milipore Sigma (or any vender)</td>
			<td>#F0291</td>
			<td></td>
		</tr>
		<tr>
			<td>Human epidermal growth factor (EGF)</td>
			<td>Milipore Sigma (or any vender)</td>
			<td>#SRP3027</td>
			<td></td>
		</tr>
		<tr>
			<td>Human meningeal cells (HMCs) isolated from human leptomeninges</td>
			<td>ScienCell</td>
			<td>#1400</td>
			<td></td>
		</tr>
		<tr>
			<td>IVIS 200 imaging system</td>
			<td>Caliper Life Sciences</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnifying glass with light</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Meningeal Cell Culture Media (MenCM)</td>
			<td>ScienCell</td>
			<td>#1401</td>
			<td></td>
		</tr>
		<tr>
			<td>Meningeal cell growth supplement (MCGS)</td>
			<td>ScienCell</td>
			<td>#1452</td>
			<td></td>
		</tr>
		<tr>
			<td>MRI imaging</td>
			<td>Bruker</td>
			<td>BioSpec series</td>
			<td>Optional</td>
		</tr>
		<tr>
			<td>P1000, P200, P20 pipettes/ pipette tips</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin-streptomycin Antibiotic solution</td>
			<td>ScienCell (or any vender)</td>
			<td>#0503</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffered saline (PBS)</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td>0.1 mm Sterile Filtered</td>
		</tr>
		<tr>
			<td>Rodent Surgical Instruments (Scissors, Forceps)</td>
			<td>Roboz Surgical Instrument (or any vendor)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Screw cap cryo tubes&#160;</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile blue paper/ drape covering</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td>n/a</td>
		</tr>
		<tr>
			<td>Sterile cotton sticks</td>
			<td>Any vendor</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue culture plates/flasks (96-well, 24-well, 12-well, 6-well, T175 etc.)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

ex vivo culture of circulating tumor cells in the cerebral spinal fluid from melanoma patients to study melanoma&#45;associated leptomeningeal disease

Leptomeningeal disease (LMD) occurs when circulating tumor cells (CTCs) disseminate into the cerebral spinal fluid (CSF) and establish in the meninges, the membrane surrounding the brain and spinal cord1,2. LMD can occur in several cancers but is particularly prevalent in melanoma. In advanced stages of melanoma, approximately 5% of patients will develop melanoma-associated M-LMD2,3. While relatively low in regard to other sites of metastasis, the consequences of M-LMD are devastating, with overall survival ranging from weeks to months, and is a significant contributor to patient morbidity1,3,4. This is primarily due to a paucity of effective treatment options combined with major gaps in our knowledge regarding how the leptomeninges are colonized by melanoma cells2. Therefore, understanding the biology of M-LMD will facilitate in designing novel therapies to improve clinical outcomes.
Recent reports have shown how CTCs colonize the unique CSF microenvironment. For example, Complement C3 promotes the invasion of tumor cells into the CSF via the choroid plexus, an intricate network of blood vessels in each ventricle of the brain5. Further, in response to the scarce micronutrients in the CSF, CTCs can upregulate lipocalin-2, an iron-scavenging protein, and its receptor SLC22A17 to enhance survival6. Using omic-based analyses of CSF, our group also found that the CSF is enriched with proteins that regulate insulin-like growth factor (IGF) signaling, as well as innate immunity3. Together, these data emphasize the value of CSF-CTCs from liquid biopsies to study M-LMD.
While CSF-CTCs can sometimes be identified by sampling patient CSF via lumbar puncture, Ommaya reservoir, or rapid autopsies, a major limitation is obtaining sufficient numbers of these rare and fragile cells1,7. For example, using the CTC enumeration technique, only several hundred to several thousand tumor cells are identifiable per patient CSF sample7, which makes it difficult to perform molecular and functional analyses in vitro or in vivo. Though there have been reports of success in briefly growing CTCs ex vivo from peripheral blood (i.e., breast cancer CTCs)8,9,10, these cells usually only grow for the short term, and there have been no reported cases of being able to grow melanoma CTCs in the CSF. Hence, finding ways to propagate melanoma CSF-CTCs, or CTCs in general, will be highly beneficial to study the biology of M-LMD7,11.
For the first time, a novel technique to propagate CSF-CTCs from M-LMD patients ex vivo is described. Here in this report, a detailed protocol was developed that allows for the culture and expansion of CSF-CTCs from M-LMD patients. Since the meninges secrete a variety of growth factors such as FGF, IGF, VEGF-A, and IGFBPs that support the growth surrounding its environment12,13,14,15,16, it was rationalized that CSF-CTCs may require these components to grow in ex vivo conditions. Therefore, this protocol uses conditioned media generated by culturing human meningeal cells- (HMCs-) in vitro. For in vivo inoculation, patient-derived cells are inoculated into immunodeficient mice to generate patient-derived CSF-CTCs (PD-CSF-CTCs) lines. The availability of patient-derived M-LMD cells will support cellular, molecular, and functional assays to study M-LMD and propose novel treatment strategies for this deadly disease.

<meta charset="utf8"></head><body>作者聚焦：评估循环肿瘤细胞在软脑膜疾病研究中的潜力

Ex Vivo （体外 ）黑色素瘤患者脑脊髓液中循环肿瘤细胞的培养以研究黑色素瘤相关的软脑膜疾病

Leptomeningeal disease from melanoma or MLMD is an aggressive form of metastasis of the meninges, where circulating tumor cells or CTCs, infiltrate into the cerebral spinal fluid. Sadly, there's no effective treatments for MLMD, so understanding its biology will help develop therapies to reduce mortality. Recent reports have shown how CTCs can colonize the nutrient scar CSF environment, such as utilizing the iron scavenging ability to enhance their survival and complement C3 to promote the invasion into the cerebral spinal fluid via the choroid plexus.Therefore, CTCs are very good tools to study MLMD. The biggest challenge in studying CSF CTCs is the overall lack of preclinical model and the scarcity of these cells. Finding ways to grow these rare and fragile cells from patients and generating a xenograft model will greatly benefit researchers in understanding MLMD pathology and designing rational therapies.We have developed a method which successfully propagated CSF CTCs from MLMD patients in-vitro and in-vivo, which provided us the ability to perform molecular and functional analyses of MLMD cells, as well as efficacy study in xenograft models. Though the current methodologies is imperfect, but being able to culture and expand CSF CTCs from melanoma patients for the first time, gave us insights on the importance of understanding the CSF Microenvironments.

Read this Scientific Article Protocol about Author Spotlight: Assessing the Potential of Circulating Tumor Cells in Leptomeningeal Disease Research at JoVE.com

Melanoma-associated leptomeningeal disease (M-LMD) occurs when circulating tumor cells (CTCs) enter into the cerebral spinal fluid (CSF) and colonize the ...

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Cancer Research

Ex Vivo Culture of Circulating Tumor Cells in the Cerebral Spinal Fluid from Melanoma Patients to Study Melanoma&#45;Associated Leptomeningeal Disease

371 Views.  University of South Florida, H. Lee Moffitt Cancer Center & Research Institute. Leptomeningeal disease from melanoma or MLMD is an aggressive form of metastasis of the meninges, where circulating tumor cells or CTCs, infiltrate into the cerebral spinal fluid. Sadly, there's no effective treatments for MLMD, so understanding its biology will help develop therapies to reduce mortality. Recent reports have shown how CTCs can colonize the nutrient scar CSF environment, such as utilizing the iron scavenging ability to enhance their survival and complement C3 to promote the in...