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Bioware® Brite Cell Line

Research Use Only. Not for use in diagnostic procedures.

Bioware® Brite Cell Line K562 Red-FLuc
Product No.: BW124735

Material Provided
Cells:

2 x 1 mL frozen aliquots (BW124735V)

Format:1.0 x 106 cells / mL in 95% FBS, 5% DMSODESIGNATION

K562 Red-FLuc

TissueSource of Parental Line
Gene Transfer Vehicle

Human: Chronic Myelogenous Leukemia (CML)
ATCC (CCL-243)
Red-FLuc-Puro 3d generation lentivirus

Bioluminescence In Vitro

At least 4,000 photons/cell/sec. Exact number will vary depending on imaging andculturing conditions.

Recommended Media andFBS

RPMI 1640 ATCC Cat. No. 30-2001.
Supplement the above with 10% Hyclone Fetal Bovine Serum (FBS) GE
HealthCare Cat. No. SH300071
Suspension cells*; viability cannot be determined solely by cell attachment.
Refer to Cell Culture Guidelines for more detailed instructions.

Culture Properties

Recommended Storage
Conditions

Remove frozen cells from dry ice packaging and immediately place cells at atemperature below -130° C, preferably in liquid nitrogen vapor, until ready to use.

Average Doubling Time15 hours

Other Recommendations

When initially thawing, use T25 flask or 10cm plate. Cells should be ready toexpand within 1-4 days. When plate is full, simply collect and dilute cells 1:3-1:7
with fresh warm media and re-plate them without using trypsin in a larger vessel.
Antibiotics can be used in the media if desired after the initial thaw. (puromycin at2ug/mL). Refer to Cell Culture Guidelines for more detailed instructions.

* Please refer to Morphology on page 2 of this document.

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Waltham, MA 02451 USAP: (800) 762-4000 or(+1) 203-925-4602
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For a complete listing of our products, visit www.perkinelmer.com.
Copyright ©2015, PerkinElmer, Inc. All rights reserved. PerkinElmer® is a registered trademark of PerkinElmer, Inc. All othertrademarks are the property of their respective owners.
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The Features
Perkin Elmer Bioware® Brite cell line models offer researchers the ability to:

Monitor early tumor development

Monitor tumor growth and metastases in vivo

Quantify tumor burden in the whole animalFollow responses to therapeutic treatments non-invasively in longitudinal studies using thesame cohorts of mice

Murine Pathogen FreeAll Perkin Elmer cell lines are confirmed to be pathogen free by the IMPACT Profile I (PCR) at theUniversity of Missouri Research Animal Diagnostic and Investigative Laboratory.
Cell Line Stability
Cell may undergo genotypic changes resulting in reduced responsiveness over time in normal cell
culture conditions. Genetic instability is a biological phenomenon that occurs in all stablytransfected cells. Therefore, it is recommended to prepare an adequate number of frozen stock atearly passages.
Product Warranty
PerkinElmer warrants that cells will be viable upon shipment from PerkinElmer for a period of thirtydays, provided they have been properly stored and handled during this period.
Human Chronic Myelogenous Leukemia (CML) Cell Line: K562 Red-FLucK562 Red-FLuc is a luciferase expressing cell line which was stably transfected with
firefly luciferase gene from Luciola Italica (Red-FLuc). The cell line was establishedby transducing lentivirus containing Red-FLuc luciferase under the control of humanubiquitin C promoter. These cells will serve as a new tool to detect drug efficacy invitro and in vivo with high sensitivity.
Morphology
K562 Red-Fluc is a suspension cell line. Cells will normally appear as rounded andfully suspended, therefore viability cannot be determined based on cell attachment.
Refer to Cell Culture Guidelines for more detailed instructions.
Bioluminescence imageof K562 Red-FLuc
subcutaneous tumor

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Growth Curve of K562 Red-FLuc Cells

Figure 1. 5 x104 cells were plated on a 6cm plate. The total numbers of cells werecounted every 24 h using a Nexcelom automatic cell counter.

In Vitro BLI Signal Stability

week 1week 2week 3week 4

Figure 2. 5 x104 cells were plated per well in 24-well plates. Cells were incubated at 37 °C for recovery overnight and luciferase assay
was performed using the PerkinElmer IVIS® SpectrumCT. Each experiment was done in quadruplicates. The cells were maintained incontinuous culture over four weeks and weekly luciferase assay was performed. Bioluminescence data was analyzed using the Living
Image 4.0 software.

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Subcutaneous Tumor Growth in a Nu/nu Mouse

day1day4

day8

day14

day18

Figure 3. 1 x 106 K562 Red-FLuc cells were injected subcutaneously into the dorsal region near the thigh of female nu/nu mouse.
Tumor growth was monitored for luciferase expression using the PerkinElmer IVIS® Spectrum at various time points. Mice wereimaged 10 minutes post i.p. injection of luciferin at 150mg/kg at various time points. The image above shows tumor growth from a
representative mouse.

Tumor Growth Comparison Between Wild Type and Red-FLuc Cells

Figure 4. 1 x 106 K562 Red-FLuc and K562 parental cells were injectedsubcutaneously into the dorsal region near the thigh of female nu/nu mouse.
Tumor growth was monitored by caliper measurements at various time points.
Similar tumor growth rate was observed for both parental and Red-FLuctransduced cell lines.

For more information on our in vivo imaging agents, please visit our website: www.perkinelmer.com/bioware.
This product is sold for in vivo animal research use only and is not intended for any diagnostic use or procedures.
Excluding purchases by authorized PerkinElmer distributors, this product is sold for use by the original purchaser
and is not for resale. Unless otherwise agreed to in writing by PerkinElmer pursuant to a separate writtenagreement, no commercial use of this product is allowed. “Commercial use” means any and all uses of this productand/or its derivatives by a party for money or other consideration and may include without limitation: (1) productmanufacture; (2) providing services, information or data to another party for remuneration; and/or (3) resale ofthe product or its derivatives, whether or not such product or derivatives are resold for use in research.
Commercial use does not include the original purchaser providing the product to its contractor solely for use on
the original purchaser’s research; provided that all product materials are returned to the original purchaser
and/or destroyed by the contractor upon completion of such project.

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