Name: Op druppels gebaseerde cytotoxiciteitstest om chimere antigeenreceptor-T-cellen op eencellig niveau te beoordelen (Video) | JoVE Journal
Uploaded: 2025-03-14T14:20:00.000+00:00
Duration: 8 min 9 s
Description: Leer een op druppels gebaseerde cytotoxiciteitstest om CAR T-cellen op eencellig niveau te beoordelen. Deze methode analyseert functionele diversiteit en verbetert het inzicht in kankerimmunotherapie.

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<ol><li>Pasqui, D. M., Latorraca, C. D. O. C., Pacheco, R. L., Riera, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((CAR-T+cell+therapy+for+patients+with+hematological+malignancies.+A+systematic+review%5BTitle%5D)+AND+%22Eur+J+Haematol%22%5BJournal%5D)">CAR-T cell therapy for patients with hematological malignancies. A systematic review.</a> Eur J Haematol. 109 (6), 601-618 (2022).</li>
<li>Krejcik, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Harnessing+the+immune+system+to+fight+multiple+myeloma%5BTitle%5D)+AND+%22Cancers%22%5BJournal%5D)">Harnessing the immune system to fight multiple myeloma.</a> Cancers. 13 (18), 4546(2021).</li>
<li>Tokarew, N., Ogonek, J., Endres, S., von Bergwelt-Baildon, M., Kobold, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Teaching+an+old+dog+new+tricks%3A+next-generation+CAR+T+cells%5BTitle%5D)+AND+%22British+J+Cancer%22%5BJournal%5D)">Teaching an old dog new tricks: next-generation CAR T cells.</a> British J Cancer. 120 (1), 26-37 (2019).</li>
<li>Bogetofte Barnkob, M., Vitting-Seerup, K., Rønn Olsen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Target+isoforms+are+an+overlooked+challenge+and+opportunity+in+chimeric+antigen+receptor+cell+therapy%5BTitle%5D)+AND+%22Immunother+Adv%22%5BJournal%5D)">Target isoforms are an overlooked challenge and opportunity in chimeric antigen receptor cell therapy.</a> Immunother Adv. 2 (1), ltac009(2022).</li>
<li>Daei Sorkhabi, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+current+landscape+of+CAR+T-cell+therapy+for+solid+tumors%3A+Mechanisms%2C+research+progress%2C+challenges%2C+and+counterstrategies%5BTitle%5D)+AND+%22Front+Immunol%22%5BJournal%5D)">The current landscape of CAR T-cell therapy for solid tumors: Mechanisms, research progress, challenges, and counterstrategies.</a> Front Immunol. 14, 1113882(2023).</li>
<li>Considerations for the Development of Chimeric Antigen Receptor (CAR) T Cell Products. , Center for Biologics Evaluation and Research. 
 <a target="_blank" href="https://www.fda.gov/regulatory-information/search-fda-guidance-documents/considerations-development-chimeric-antigen-receptor-car-t-cell-products">https://www.fda.gov/regulatory-information/search-fda-guidance-documents/considerations-development-chimeric-antigen-receptor-car-t-cell-products</a> (2024).</li>
<li>Roddie, C., O'Reilly, M., Dias Alves Pinto, J., Vispute, K., Lowdell, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Manufacturing+chimeric+antigen+receptor+T+cells%3A+issues+and+challenges%5BTitle%5D)+AND+%22Cytotherapy%22%5BJournal%5D)">Manufacturing chimeric antigen receptor T cells: issues and challenges.</a> Cytotherapy. 21 (3), 327-340 (2019).</li>
<li>Wang, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Improvement+of+in+vitro+potency+assays+by+a+resting+step+for+clinical-grade+chimeric+antigen+receptor+engineered+T+cells%5BTitle%5D)+AND+%22Cytotherapy%22%5BJournal%5D)">Improvement of in vitro potency assays by a resting step for clinical-grade chimeric antigen receptor engineered T cells.</a> Cytotherapy. 21 (5), 566-578 (2019).</li>
<li>Burrows, S. R., Fernan, A., Argaet, V., Suhrbier, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bystander+apoptosis+induced+by+CD8%2B+cytotoxic+T+cell+%28CTL%29+clones%3A+implications+for+CTL+lytic+mechanisms%5BTitle%5D)+AND+%22Int+Immunol%22%5BJournal%5D)">Bystander apoptosis induced by CD8+ cytotoxic T cell (CTL) clones: implications for CTL lytic mechanisms.</a> Int Immunol. 5 (9), 1049-1058 (1993).</li>
<li>Gattinoni, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Acquisition+of+full+effector+function+in+vitro+paradoxically+impairs+the+in+vivo+antitumor+efficacy+of+adoptively+transferred+CD8%2B+T+cells%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells.</a> J Clin Invest. 115 (6), 1616-1626 (2005).</li>
<li>Chen, G. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Integrative+bulk+and+single-cell+profiling+of+premanufacture+T-cell+populations+reveals+factors+mediating+long-term+persistence+of+CAR+T-cell+therapy%5BTitle%5D)+AND+%22Cancer+Dis%22%5BJournal%5D)">Integrative bulk and single-cell profiling of premanufacture T-cell populations reveals factors mediating long-term persistence of CAR T-cell therapy.</a> Cancer Dis. 11 (9), 2186-2199 (2021).</li>
<li>Frazer, G. L., Gawden-Bone, C. M., Dieckmann, N. M. G., Asano, Y., Griffiths, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Signal+strength+controls+the+rate+of+polarization+within+CTLs+during+killing%5BTitle%5D)+AND+%22J+Cell+Biol%22%5BJournal%5D)">Signal strength controls the rate of polarization within CTLs during killing.</a> J Cell Biol. 220 (10), e202104093(2021).</li>
<li>Hermans, I. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+VITAL+assay%3A+a+versatile+fluorometric+technique+for+assessing+CTL-+and+NKT-mediated+cytotoxicity+against+multiple+targets+in+vitro+and+in+vivo%5BTitle%5D)+AND+%22J+Immunol+Meth%22%5BJournal%5D)">The VITAL assay: a versatile fluorometric technique for assessing CTL- and NKT-mediated cytotoxicity against multiple targets in vitro and in vivo.</a> J Immunol Meth. 285 (1), 25-40 (2004).</li>
<li>Zaritskaya, L., Shurin, M. R., Sayers, T. J., Malyguine, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+flow+cytometric+assays+for+monitoring+cell-mediated+cytotoxicity%5BTitle%5D)+AND+%22Expert+Rev+Vaccines%22%5BJournal%5D)">New flow cytometric assays for monitoring cell-mediated cytotoxicity.</a> Expert Rev Vaccines. 9 (6), 601-616 (2010).</li>
<li>Piccinini, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((In+vitro+CAR-T+cell+killing%3A+validation+of+the+potency+assay%5BTitle%5D)+AND+%22Cancer+Immunol+Immunother+CII%22%5BJournal%5D)">In vitro CAR-T cell killing: validation of the potency assay.</a> Cancer Immunol Immunother CII. 73 (9), 168(2024).</li>
<li>Martinez, E. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High-throughput+flow+cytometric+method+for+the+simultaneous+measurement+of+CAR-T+cell+characterization+and+cytotoxicity+against+solid+tumor+cell+lines%5BTitle%5D)+AND+%22SLAS+Disc+Adv+Life+Sci%22%5BJournal%5D)">High-throughput flow cytometric method for the simultaneous measurement of CAR-T cell characterization and cytotoxicity against solid tumor cell lines.</a> SLAS Disc Adv Life Sci. 23 (7), 603-612 (2018).</li>
<li>Jedema, I., van der Werff, N. M., Barge, R. M. Y., Willemze, R., Falkenburg, J. H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+CFSE-based+assay+to+determine+susceptibility+to+lysis+by+cytotoxic+T+cells+of+leukemic+precursor+cells+within+a+heterogeneous+target+cell+population%5BTitle%5D)+AND+%22Blood%22%5BJournal%5D)">New CFSE-based assay to determine susceptibility to lysis by cytotoxic T cells of leukemic precursor cells within a heterogeneous target cell population.</a> Blood. 103 (7), 2677-2682 (2004).</li>
<li>Liu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cellular+and+molecular+imaging+of+CAR-T+cell-based+immunotherapy%5BTitle%5D)+AND+%22Adv+Drug+Delivery+Rev%22%5BJournal%5D)">Cellular and molecular imaging of CAR-T cell-based immunotherapy.</a> Adv Drug Delivery Rev. 203, 115135(2023).</li>
<li>Liadi, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Defining+potency+of+CAR%2B+T+cells%3A+Fast+and+furious+or+slow+and+steady%5BTitle%5D)+AND+%22Oncoimmunology%22%5BJournal%5D)">Defining potency of CAR+ T cells: Fast and furious or slow and steady.</a> Oncoimmunology. 8 (10), e1051298(2019).</li>
<li>Xhangolli, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Single-cell+analysis+of+CAR-T+cell+activation+reveals+a+mixed+TH1%2FTH2+response+independent+of+differentiation%5BTitle%5D)+AND+%22Genom+Proteom+Bioinfo%22%5BJournal%5D)">Single-cell analysis of CAR-T cell activation reveals a mixed TH1/TH2 response independent of differentiation.</a> Genom Proteom Bioinfo. 17 (2), 129-139 (2019).</li>
<li>Zhou, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Evaluation+of+single-cell+cytokine+secretion+and+cell-cell+interactions+with+a+hierarchical+loading+microwell+chip%5BTitle%5D)+AND+%22Cell+Rep%22%5BJournal%5D)">Evaluation of single-cell cytokine secretion and cell-cell interactions with a hierarchical loading microwell chip.</a> Cell Rep. 31 (4), 107574(2020).</li>
<li>LaBelle, C. A., Zhang, R. J., Hunsucker, S. A., Armistead, P. M., Allbritton, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Microraft+arrays+for+serial-killer+CD19+chimeric+antigen+receptor+T+cells+and+single+cell+isolation%5BTitle%5D)+AND+%22Cytometry+J+Int+Soc+Anal+Cytol%22%5BJournal%5D)">Microraft arrays for serial-killer CD19 chimeric antigen receptor T cells and single cell isolation.</a> Cytometry J Int Soc Anal Cytol. 103 (3), 208-220 (2023).</li>
<li>Hellmann, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Heterogeneously+deacetylated+chitosans+possess+an+unexpected+regular+pattern+favoring+acetylation+at+every+third+position%5BTitle%5D)+AND+%22Nat+Comm%22%5BJournal%5D)">Heterogeneously deacetylated chitosans possess an unexpected regular pattern favoring acetylation at every third position.</a> Nat Comm. 15 (1), 6695(2024).</li>
<li>Wong, K. U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Assessment+of+chimeric+antigen+receptor+T+cytotoxicity+by+droplet+microfluidics+in+vitro%5BTitle%5D)+AND+%22Antibody+Therapeut%22%5BJournal%5D)">Assessment of chimeric antigen receptor T cytotoxicity by droplet microfluidics in vitro.</a> Antibody Therapeut. 5 (2), 85-99 (2022).</li>
<li>Antona, S., Platzman, I., Spatz, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Droplet-based+cytotoxicity+assay%3A+Implementation+of+time-efficient+screening+of+antitumor+activity+of+natural+killer+cells%5BTitle%5D)+AND+%22ACS+omega%22%5BJournal%5D)">Droplet-based cytotoxicity assay: Implementation of time-efficient screening of antitumor activity of natural killer cells.</a> ACS omega. 5 (38), 24674-24683 (2020).</li>
<li>Subedi, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+automated+real-time+microfluidic+platform+to+probe+single+NK+cell+heterogeneity+and+cytotoxicity+on-chip%5BTitle%5D)+AND+%22Sci+Rep%22%5BJournal%5D)">An automated real-time microfluidic platform to probe single NK cell heterogeneity and cytotoxicity on-chip.</a> Sci Rep. 11 (1), 17084(2021).</li>
<li>Zhang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Enhanced+CRISPR%2FCas12a-based+quantitative+detection+of+nucleic+acids+using+double+emulsion+droplets%5BTitle%5D)+AND+%22Biosens+Bioelect%22%5BJournal%5D)">Enhanced CRISPR/Cas12a-based quantitative detection of nucleic acids using double emulsion droplets.</a> Biosens Bioelect. 257, 116339(2024).</li>
<li>Kochenderfer, J. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Construction+and+preclinical+evaluation+of+an+anti-CD19+chimeric+antigen+receptor%5BTitle%5D)+AND+%22J+Immunother%22%5BJournal%5D)">Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor.</a> J Immunother. 32 (7), 689-702 (2009).</li>
<li>Michaels, Y. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Precise+tuning+of+gene+expression+levels+in+mammalian+cells%5BTitle%5D)+AND+%22Nat+Comm%22%5BJournal%5D)">Precise tuning of gene expression levels in mammalian cells.</a> Nat Comm. 10 (1), 818(2019).</li>
<li>Yuan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Droplet+encapsulation+improves+accuracy+of+immune+cell+cytokine+capture+assays%5BTitle%5D)+AND+%22Lab+on+a+Chip%22%5BJournal%5D)">Droplet encapsulation improves accuracy of immune cell cytokine capture assays.</a> Lab on a Chip. 20 (8), 1513-1520 (2020).</li>
<li>Brower, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Double+emulsion+flow+cytometry+with+high-throughput+single+droplet+isolation+and+nucleic+acid+recovery%5BTitle%5D)+AND+%22Lab+on+a+Chip%22%5BJournal%5D)">Double emulsion flow cytometry with high-throughput single droplet isolation and nucleic acid recovery.</a> Lab on a Chip. 20 (12), 2062-2074 (2020).</li>
<li>Kirouac, D. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Deconvolution+of+clinical+variance+in+CAR-T+cell+pharmacology+and+response%5BTitle%5D)+AND+%22Nat+Biotechnol%22%5BJournal%5D)">Deconvolution of clinical variance in CAR-T cell pharmacology and response.</a> Nat Biotechnol. 41 (11), 1606-1617 (2023).</li>
</ol>

<table><tbody><tr><td>Reagents</td><td></td><td></td><td></td></tr><tr><td>Blocker BSA (BSA)</td><td>Thermo Scientific</td><td>37525</td><td></td></tr><tr><td>CellTrace Far Red Cell Proliferation Kit (far red fluorescent dye)</td><td>Invitrogen</td><td>C34564</td><td></td></tr><tr><td>CellTrace Violet Cell Proliferation Kit (violet fluorescent dye)</td><td>Invitrogen</td><td>C34557</td><td></td></tr><tr><td>DE Stabilizing Solution (stabilizing solution)</td><td>Samplix</td><td>REDIVSTABSOL1500</td><td></td></tr><tr><td>DPBS (dPBS)</td><td>Gibco</td><td>14190-094</td><td></td></tr><tr><td>Dulbecco&rsquo;s PBS (dPBS)</td><td>Capricorn Scientific</td><td>PBS-1A</td><td></td></tr><tr><td>Dynabeads Human T-Activator (CD3/CD28 activation beads)</td><td>Gibco</td><td>11132D</td><td></td></tr><tr><td>Fetal Bovine Serum (FBS)</td><td>Capricorn Scientific</td><td>FBS-HI-12A</td><td></td></tr><tr><td>Lenti-X Concentrator (PEG-based reagent)</td><td>Takara Bio</td><td>631232</td><td></td></tr><tr><td>MACSelect LNGFR Microbeads (anti-NGFR magnetic beads)</td><td>Miltenyi Biotec</td><td>130-091-330</td><td></td></tr><tr><td>OptiPrep density gradient medium (gradient medium)</td><td>Stemcell</td><td>7820</td><td></td></tr><tr><td>Penicillin-Streptomycin (P/S)</td><td>Capricorn Scientific</td><td>PS-B</td><td></td></tr><tr><td>Propidium iodide (PI)</td><td>Invitrogen</td><td>BMS500PI</td><td></td></tr><tr><td>Recombinant Human IL-2 (IL-2)</td><td>Peprotech</td><td>200-02</td><td></td></tr><tr><td>RPMI-1640 with Stable Glutamine (RPMI-1640)</td><td>Capricorn Scientific</td><td>RPMI-STA</td><td></td></tr><tr><td>RPMI-1640 without L-Glutamine and phenol red</td><td>Gibco</td><td>32404-014</td><td></td></tr><tr><td>Xdrop DE oil I (oil)</td><td>Samplix</td><td>REOILDEC1900</td><td></td></tr><tr><td>Xdrop Granzyme B substrate (GzmB substrate)</td><td>Samplix</td><td>REGRB100</td><td></td></tr><tr><td>Zombie-NIR viability dye (viability dye)</td><td>BioLegend</td><td>423106</td><td></td></tr><tr><td>Plasticware etc.</td><td></td><td></td><td></td></tr><tr><td>8-chamber glass slide</td><td>Chemometec</td><td>942-0003</td><td></td></tr><tr><td>Cell culture plate, 12 well</td><td>TH Geyer</td><td>7696791</td><td></td></tr><tr><td>DNA LoBind tube, 2 mL (DNA tube)</td><td>Eppendorf</td><td>30108078</td><td></td></tr><tr><td>Eppendorf tube, 1.5 mL (1.5 mL tube)</td><td>Eppendorf</td><td>30108051</td><td></td></tr><tr><td>Falcon tube, 15 mL (15 mL tube)</td><td>TPP</td><td>91015</td><td></td></tr><tr><td>Falcon tube, 5 mL (5 mL tube)</td><td>Falcon (VWR)</td><td>734-0443</td><td></td></tr><tr><td>Green cell suspension flasks for cell incubations (T75 flask)</td><td>Sarstedt</td><td>148.19.22</td><td></td></tr><tr><td>Green cell suspension plates for cell incubations (96 well plate)</td><td>Sarstedt</td><td>148.32.20</td><td></td></tr><tr><td>LS Separation Columns (separation column)</td><td>Miltenyi Biotec</td><td>130-042-401</td><td></td></tr><tr><td>Xdrop DE Gaskets (gaskets)</td><td>Samplix</td><td>#GADEA100</td><td></td></tr><tr><td>Xdrop DE50 Cartridge (encapsulation cartridge)</td><td>Samplix</td><td>#CADE50A100</td><td></td></tr><tr><td>Antibodies</td><td></td><td></td><td></td></tr><tr><td>anti-CCR7 PE-Dazzle 594</td><td>BioLegend</td><td>353236</td><td></td></tr><tr><td>anti-CD19 CAR FMC63 Idiotype Antibody, PE</td><td>Miltenyi Biotec</td><td>130-127-342</td><td></td></tr><tr><td>anti-CD3 APC</td><td>Biolegend</td><td>300439</td><td></td></tr><tr><td>anti-CD3 BV480</td><td>BD Biosciences</td><td>566105</td><td></td></tr><tr><td>anti-CD3 FITC</td><td>BD Biosciences</td><td>345763</td><td></td></tr><tr><td>anti-CD4 BUV661</td><td>BD Biosciences</td><td>612962</td><td></td></tr><tr><td>anti-CD4 StarBright Violet 760</td><td>Bio-Rad</td><td>MCA1267SBV760T</td><td></td></tr><tr><td>anti-CD45RA BUV395</td><td>BD Biosciences</td><td>740298</td><td></td></tr><tr><td>anti-CD8 PE-Cy7</td><td>BioLegend</td><td>344712</td><td></td></tr><tr><td>anti-CD8 StarBright Violet</td><td>Bio-Rad</td><td>MCA1226SBV760</td><td></td></tr><tr><td>anti-NGFR FITC</td><td>BioLegend</td><td>345106</td><td></td></tr><tr><td>anti-NGFR PE</td><td>BioLegend</td><td>345106</td><td></td></tr><tr><td>Cells</td><td></td><td></td><td></td></tr><tr><td>JeKo-1 Mantle-cell lymphoma cell-line (JeKo-1)</td><td>ATCC</td><td>CRL-3006</td><td></td></tr><tr><td>Primary peripheral blood mononuclear cells (PBMCs)</td><td></td><td></td><td></td></tr><tr><td>Equipment</td><td></td><td></td><td></td></tr><tr><td>Countess 3 Automated Cell Counter</td><td>Thermo Fisher Scientific</td><td>A50298</td><td></td></tr><tr><td>DynaMag-2 Magnet</td><td>Invitrogen</td><td>12321D</td><td></td></tr><tr><td>NovoCyte Quanteon Flow Cytometer (flow cytometer)</td><td>Agilent</td><td>2010011AA</td><td></td></tr><tr><td>Xdrop (microfluidics device)</td><td>Samplix</td><td>IN00110-EU</td><td></td></tr></tbody></table>

op druppels gebaseerde cytotoxiciteitstest om chimere antigeenreceptor-t-cellen op eencellig niveau te beoordelen

Droplet-based Cytotoxicity Assay to Assess Chimeric Antigen Receptor T cells at the Single-cell Level

Op druppels gebaseerde cytotoxiciteitstest om chimere antigeenreceptor-T-cellen op eencellig niveau te beoordelen

The assessment of the cytotoxic potential of T cell-based therapies, such as chimeric antigen receptor (CAR) T cell treatments, is instrumental in ...

droplet-based-cytotoxicity-assay-to-assess-chimeric-antigen-receptor

Research

JoVE Journal

Immunology and Infection

Immunologie en Infectie

This video demonstrates a method for monitoring CAR T cell activity against cancer cells through real-time electrical impedance measurements. Tracking changes in impedance assesses the effectiveness of CAR T cells in inducing cancer cell cytolysis, providing insights into their cytotoxic capabilities.

A Real-Time In Vitro Cytolysis Assay to Evaluate the Potency of CAR T Cells

We beschrijven een single-cell high-throughput assay voor cytotoxiciteit van T-cellen te meten wanneer geïncubeerd met tumor doelcellen. Deze werkwijze gebruikt een dichte, elastomeer reeks sub-nanoliter wells (~ 100.000 putjes / array) ruimtelijk beperken de T-cellen en doelcellen met gedefinieerde verhoudingen en is gekoppeld met fluorescentie microscopie effector-target conjugatie en daaropvolgende apoptose controleren.

Quantitative High-throughput Eencellige cytotoxiciteit T cellen

We beschrijven een kwantitatieve real-time in vitro cytolysis assay systeem om de potentie van chimere antigeen receptor T-cellen gericht op vloeibare en solide tumorcellen te evalueren. Dit protocol kan worden uitgebreid om andere immuuneffector cellen te beoordelen, evenals combinatie behandelingen.

Een real-time potentie test voor Chimerische antigeen receptor T-cellen gericht op vaste en hematologische kankercellen

Hier beschrijven we een in vitro co cultuur methode recursief uitdaging tumor-gerichte T-cellen, die het mogelijk voor fenotypische en functionele analyse van antitumorale T cel activiteit maakt.

In Vitro Tumor cel Rechallenge voor voorspellende evaluatie van chimeer antigeen Receptor T cel antitumorale functie

Hier beschrijven we een protocol waarin een acuut lymfoblastische leukemiepatiënt-afgeleid xenograftmodel wordt gebruikt als een strategie om CD19-gerichte chimere antigeenreceptor T-cel-geassocieerde toxiciteiten te beoordelen en te monitoren.

Beoordeling van chimere antigeenreceptor T-celgeassocieerde toxiciteiten met behulp van een acuut lymfoblastische leukemiepatiënt-afgeleid xenograftmuismodel

Een protocol om kwantitatieve tumorceldoding te evalueren door Jurkat-cellen die chimere antigeenreceptor (CAR) tot expressie brengen die gericht zijn op een enkel tumorantigeen. Dit protocol kan worden gebruikt als een screeningsplatform voor snelle optimalisatie van CAR-scharnierconstructies voorafgaand aan bevestiging in perifere bloed-afgeleide T-cellen.

Snelle in-vitrocytotoxiciteitsevaluatie van Jurkat die chimere antigeenreceptor tot expressie brengt met behulp van fluorescerende beeldvorming

Dit protocol schetst een gedetailleerde procedure voor het genereren van voorbijgaande chimere antigeenreceptor (CAR) T-cellen met behulp van niet-integrerend mRNA voor kankerimmunotherapie en biedt betrouwbare methoden voor het evalueren van CAR-T-cellen en hun cytotoxische functie.

Een niet-virale benadering om voorbijgaande chimere antigeenreceptor-T-cellen te genereren met behulp van mRNA voor immunotherapie bij kanker

Dit protocol biedt een gestandaardiseerde in vitro. workflow voor consistente meting van natuurlijke, chimere antigeenreceptor-gemedieerde en antistofafhankelijke cytotoxiciteit van menselijke natuurlijke moordenaars met behulp van live-cell imaging en flowcytometrie.

Kwantificatie van NK-cel cytotoxiciteit versterkt door chimerische antigeenreceptoren en therapeutische antilichamen met behulp van live-cell imaging en flowcytometrie

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HIV-1-pathogenese wordt bepaald door zowel virale kenmerken als genetische factoren van de gastheer. Hier beschrijven we een robuuste methode die reproduceerbare metingen mogelijk maakt om de impact van de gag gensequentieverschil op de in vitro replicatiecapaciteit van het virus te beoordelen.

Een restrictie-enzym Based Klonen Methode om de Beoordelen<em&gt; In vitro</em&gt; Replicatie Capaciteit van HIV-1 Subtype C Gag-MJ4 Chimeric Virussen

Leer een op druppels gebaseerde cytotoxiciteitstest om CAR T-cellen op eencellig niveau te beoordelen. Deze methode analyseert functionele diversiteit en verbetert het inzicht in kankerimmunotherapie.

Op druppels gebaseerde cytotoxiciteitstest om chimere antigeenreceptor-T-cellen op eencellig niveau te beoordelen

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