{"product_id":"cl183","title":"Nalm6-Fluc-Puro\/HuCD19-Puro (mid)","description":"\u003cdiv style=\"mso-element: comment-list;\"\u003e\n\u003cp\u003eThe Nalm6-Fluc-Puro\/HuCD19-Puro (mid) cell line stably expresses firefly luciferase and levels of human CD19 that are below the endogenous levels normally observed in Nalm6 cells (see Characterization tab for data). The cells are designed for \u003cem\u003ein vitro \u003c\/em\u003eand \u003cem\u003ein vivo \u003c\/em\u003estudies of tumor cells expressing CD19. These cells can be used in combination with other Imanis Nalm6-Fluc-Puro cell lines that are part of the CD19 panel comprised of Nalm6-Fluc-Puro cells with various CD19 surface receptor densities:\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/imanislife.com\/products\/nalm6-fluc-puro\/\"\u003eNalm6-Fluc-Puro (endogenous (high) levels of CD19)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/imanislife.com\/products\/nalm6-fluc-puro-cd19-ko\/\"\u003eNalm6-Fluc-Puro\/CD19-KO (negative for CD19)\u003c\/a\u003e\u003c\/li\u003e\n\u003cli\u003e\u003ca href=\"https:\/\/imanislife.com\/?post_type=products\u0026amp;p=4275\"\u003eNalm6-Fluc-Puro\/HuCD19-Puro (low) (moderate levels of CD19)\u003c\/a\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eTo generate this cell line, \u003ca href=\"https:\/\/imanislife.com\/products\/nalm6-fluc-puro-cd19-ko\/\"\u003eNalm6-Fluc-Puro\/CD19-KO\u003c\/a\u003e cells were transduced with a human CD19-encoding lentivirus. A moderate HuCD19 expressing population was generated by selection using a methylcellulose based semi-solid medium.\u003c\/p\u003e\n\u003cp\u003eThis cell line has been tested for mycoplasma contamination and is certified mycoplasma free.\u003c\/p\u003e\n\u003cp\u003eThe parental Nalm6 cell line was licensed and purchased directly from ATCC*.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eReplication Competent Lentivirus (RCL) Test \u003c\/strong\u003e(including a test report) is available for this cell line at an added cost. \u003ca href=\"https:\/\/imanislife.com\/contact-us\/\"\u003e\u003cstrong\u003eContact us\u003c\/strong\u003e\u003c\/a\u003e to learn more.\u003c\/p\u003e\n\u003cp\u003e*The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eCharacterization\u003c\/strong\u003e\u003c\/p\u003e\n\u003ch3\u003eMorphology\u003c\/h3\u003e\n\u003chr\u003e\n\u003cp\u003e\u003cimg class=\"alignright size-medium wp-image-4285\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/CL183-Morphology-400x302.jpg\" alt=\"\"\u003ePhase cell photo taken at 200x magnification\u003c\/p\u003e\n\u003ch3\u003eLuciferase Expression\u003c\/h3\u003e\n\u003chr\u003e\n\u003cp\u003e\u003cimg class=\"alignright size-full wp-image-4286\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/Luc-Exp-CL183.png\" alt=\"\"\u003e\u003c\/p\u003e\n\u003ch3\u003eCD19 Expression\u003c\/h3\u003e\n\u003chr\u003e\n\u003cdiv\u003e\u003cimg class=\"alignright size-medium wp-image-4287\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/CL183-CD19-400x236.png\" alt=\"\"\u003e\u003c\/div\u003e\n\u003cp\u003eNalm6-Fluc-Puro\/HuCD19-Puro (mid) cells were stained with an anti-CD19 (blue) or isotype control (grey) antibody and analyzed by flow cytometry.\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cimg class=\"alignright size-medium wp-image-4288\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/CL183-400x165.png\" alt=\"\"\u003eThe indicated cell lines were stained with an anti-CD19 (blue) or isotype control (grey) antibody and analyzed by flow cytometry. The antibody binding capacity, representing the number of CD19 receptors on the surface of the cells was quantitated using a Quantum-Simply cellular kit.\u003c\/p\u003e\n\u003ch3\u003eSurface Receptor Profiling\u003c\/h3\u003e\n\u003chr\u003e\n\u003cp\u003e\u003cimg class=\"alignright size-medium wp-image-4289\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/Surface-Receptor-CL183-400x306.png\" alt=\"\"\u003eNalm6-Fluc-Puro\/HuCD19-Puro (mid) cells (blue) were stained with anti-CD20 (A), anti-BCMA (B), anti-GPRC5D (C), anti-CD38 (D), or isotype control (grey; all panels) antibody and analyzed by flow cytometry.\u003c\/p\u003e\n\u003ch3\u003eCAR-T Activation Assay\u003c\/h3\u003e\n\u003chr\u003e\n\u003cp\u003e\u003cimg class=\"alignright size-medium wp-image-4290\" src=\"https:\/\/imanislife.com\/wp-content\/uploads\/2024\/11\/CAR-t-CL183-400x163.png\" alt=\"\"\u003eJurkat T cells expressing a CD19-CAR and GFP were generated by lentivector transduction. Tranduced or untransduced parental Jurkat were incubated in a 1:1 ratio with the indicated Nalm6 cell lines and the following day flow cytometry was performed. The CD3+ Jurkat cells were analyzed for CD19CAR-GFP expression and CD69 expression, which is upregulated in activated T cells.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eGrowth Conditions\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eComplete Growth Medium: RPMI-1640 (IMDM) supplemented with 10% fetal bovine serum (FBS), 10 mM HEPES, and 1% Penicillin\/Streptomycin.\u003cbr\u003eFor long-term maintenance (more than two to three weeks) addition of 1 µg\/mL of puromycin to the media is recommended to maintain high Fluc and HuCD19 expression.\u003cbr\u003eThese cells should be grown in the indicated medium and subcultured as needed to maintain a density between 5 x 105 and 3 x 106 cells\/mL. The cells can be passaged by dilution in fresh medium, with occasional passaging using centrifugation to limit the amount of debris in cultures.\u003c\/p\u003e\n\u003cp\u003e \u003cstrong\u003eUsage Information\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003eThese cells are suitable for \u003cem\u003ein vitro\u003c\/em\u003e and \u003cem\u003ein vivo\u003c\/em\u003e experimentation. The Fluc transgene facilitates \u003cem\u003ein vivo\u003c\/em\u003e noninvasive bioluminescent imaging of implanted cells and quantitation of cells\u003cspan\u003e \u003c\/span\u003e\u003cem\u003ein vitro\u003c\/em\u003e.\u003c\/p\u003e\n\u003cp\u003eThese cells were generated via lentiviral vector transduction. The lentiviral vectors used for transduction were self-inactivating (SIN) vectors in which the viral enhancer and promoter have been deleted. Transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization by replication competent viruses and enables regulated expression of the genes from the internal promoters without \u003cem\u003ecis\u003c\/em\u003e-acting effects of the LTR\u003csup\u003e1\u003c\/sup\u003e. Nevertheless, all work with these cells should be performed under biosafety-level 2 (BSL2) conditions by trained personnel. Institutional requirements may permit handling of these cells under BSL1 conditions if certain criteria are met.\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003eMiyoshi, H\u003cspan\u003e \u003c\/span\u003e\u003cem\u003eet al.\u003cspan\u003e \u003c\/span\u003e\u003c\/em\u003e(1998). Development of a self-inactivating lentivirus vector.\u003cspan\u003e \u003c\/span\u003e\u003cem\u003eJournal of Virology\u003c\/em\u003e\u003cspan\u003e \u003c\/span\u003e72: 8150-8157.\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003chr class=\"msocomoff\" align=\"left\" size=\"1\" width=\"33%\"\u003e\n\u003c!--[endif]--\u003e\n\u003cdiv style=\"mso-element: comment;\"\u003e\n\u003c!-- [if !supportAnnotations]--\u003e\n\u003cdiv id=\"_com_1\" class=\"msocomtxt\" language=\"JavaScript\" onmouseover=\"msoCommentShow('_anchor_1','_com_1')\" onmouseout=\"msoCommentHide('_com_1')\"\u003e\n\u003c!--[endif]--\u003e\u003cspan style=\"mso-comment-author: 'Rianna Vandergaast'; mso-comment-providerid: AD; mso-comment-userid: 'S::vandergaast\\.rianna\\@imanislife\\.com::e78267c4-30da-4740-afd1-63997f420c96';\"\u003e\u003c!-- [if !supportAnnotations]--\u003e\u003ca name=\"_msocom_1\"\u003e\u003c\/a\u003e\u003c!--[endif]--\u003e\u003c\/span\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cspan class=\"MsoCommentReference\"\u003e\u003cspan style=\"font-size: 8.0pt;\"\u003e\u003cspan style=\"mso-special-character: comment;\"\u003e\u003c\/span\u003e\u003c\/span\u003e\u003c\/span\u003e\n\u003c\/div\u003e","brand":"Imanis Life Sciences","offers":[{"title":"Frozen \/ Standard","offer_id":44811841405109,"sku":"CL183","price":3500.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/imanis-life-sciences-4.myshopify.com\/products\/cl183","provider":"Imanis Life Sciences","version":"1.0","type":"link"}