Mit Luciferase markierte Zelllinien: ein wertvolles Tool für Onkologiestudien

Author: Manali Dimri, PhD | Scientist, Scientific Development
Date: July 2021


Luciferases are oxidative enzymes that emit light in the presence of a substrate (D-luciferin) within a living organism, a process known as bioluminescence. The gene for the most common luciferases comes from the family of light producing enzymes called firefly luciferases1, 2

In preclinical research, small animal in vivo imaging plays an essential role in visualizing physiological processes, progression of disease and development of therapies. Luciferase (luc) enabled cell lines offer a simple, high-throughput and robust means to quantitatively assess tumor burden and response of tumors to treatment therapies in subject animals, through bioluminescence imaging (BLI)1, 2. Click here to learn more about BLI

The sensitivity and accuracy of BLI systems offers multiple advantages over traditional methods, such as:

(1)    Noninvasive real-time whole-body in vivo tumor monitoring and imaging

(2)    Continuous assessment of tumor progression and response to therapeutic treatments in the same animal

(3)    Metastasis assessment

(4)    Reduced need for animal sacrifice

Why Labcorp Drug Development (formerly Covance Laboratories)?

Covance was the first Contract Research Organization (CRO) to offer BLI, in 2003, and in the past 18 years we have accrued extensive experience in this optical imaging field. We offer a large panel of luciferase-enabled cell lines with over 80 unique hematological malignancy cell lines (Table 1). We have a dedicated team of experts to design the best oncology study for you as well as to ensure smooth study execution. Our scientists are skilled to engineer our in-house cell lines or your cell line of interest as well as to custom make vectors to express luciferase for BLI detection. Complete list of cell lines

LabCorp Drug Development (formerly Covance Laboratories) has a license agreement from Dana Farber Cancer Institute and other organizations that provides additional access to many characterized, in vivo validated luciferase-expressing tumor lines.

Our luciferase-expressing tumor cell lines have

·       Stable luciferase expression

·       A fluorescent protein (mCherry) along with the Luc 2 gene

·       Quantitative correlation between signal strength and cell numbers

·       High sensitivity and low signal-to-noise ratio

·       Availability of multiple tumor cell lines from human, mouse, and rat

·       Suitable for in vitro as well as for in vivo assays

We also offer an alternative method for cell transduction in luc- cell enabling which is cell electroporation allowing clients to choose customized vectors instead of a lentiviral vector.

Service offerings related to BLI include:

How it works?

Typically, cancer cells are engineered to express the firefly luciferase gene along with a puromycin resistance gene using a lentiviral system for transduction (Fig. 1A). Along with the Luc 2 gene, the construct has a fluorescent protein coding gene (mCherry) that enables the detection of tumor cells in different tissues over time. Cells are cultured in the presence of puromycin to select the cells with inserted lentiviral vector encoding firefly luciferase (Fig. 1B). Light output is generated (Fig. 1C(ii)) from the luciferase enabled cells and bioluminescence is measured using the IVIS® In Vivo Imaging System (Fig. 1C (i)); Luc-enabled cells are then engrafted into mice to form tumors. Following the injection of D-luciferin (substrate), the luciferase enzyme will catalyze this substrate resulting in light emission detected with IVIS® (PerkinElmer, Waltham, MA) (Fig. 1D(i)) and analyzed in regions of interest using the Perkin Elmer's Living Image software (Fig. 1D (ii)).

Fig. 1: Representation of generation of luciferase-expressing tumor cell lines for in vivo bioluminescence imaging. A. Tumor cell lines are transduced with a lentivirus vector. B. Transduced tumor cell lines are selected and expanded using selection marker. C. (i) Raji cells were transduced with lentiviral vector and luciferase expression was confirmed upon addition of D-luciferin substrate. (ii) Luciferase bioluminescence was quantified using Perkin Elmer's Living Image software and number of cells vs total flux (p/s= photons/second) was plotted. D. (i) Monitoring tumor burden using BLI. Luminescence imaged using IVIS® at different time points. (ii) Luciferase bioluminescence was quantified using the Perkin Elmer's Living Image software.

Histotyp

Cell line

Art

Gehirn

D54-Luc

Mensch

 

Gli36-DsRed-R-Luc (gerettet)

Mensch

 

LN-827(pMMP-LucNeo)

Mensch

 

U-251-Luc-mCh-Puro

Mensch

 

U-87 MG-Luc

Mensch

 

GL261-Luc2

Maus

 

9L-Luc

Ratte

Blase

T24-Luc-Neo

Mensch

 

MB49-Luc-mCh-Puro

Maus

Dickdarm

COLO 205-Luc #2

Mensch

 

HCT-116-Luc

Mensch

 

HT-29-Luc

Mensch

 

CT26.WT-luc-mCh-puro

Maus

 

MC38-NCI.TD1-luc-mCh-puro

Maus

Endometrial

KLE-Luc-mCh-Puro

Mensch

Leukämie [AML]

Kasumi-3-Luc-mCh-Puro

Mensch

 

KG-1-Luc-mCh-Puro

Mensch

 

MV-4-11-Luc-mCh-Puro

Mensch

 

C1498-Luc-mCh-Puro

Maus

Leukämie [B-ALL]

NALM6-Luc-MCh-Puro

Mensch

 

Reh (pMMP-Luc-Neo)

Mensch

Leukämie [CML]

K-562-Luc2

Mensch

Leukemia [erythro]

HEL 92.1.7-Luc-Neo

Mensch

 

HEL-Luc-Neo

Mensch

Leukämie [T-ALL]

DND-41-Luc-mCh-Puro

Mensch

 

MOLT-4-Luc-MCh-Puro

Mensch

Leber

BNL 1ME A.7R.1-Luc-mCh-Puro

Maus

 

Hep-55,1C-Luc-mCh-Puro

Maus

 

Hepa 1-6-Luc-mCh-Puro

Maus

Lunge

LL/2-Luc-M38

Maus

 

AB1-Luc-mCh-puro

Maus

Lunge [NSCLC]

A549-Luc-C8

Mensch

 

HCC827-Luc-mCh-Puro

Mensch

 

NCI-H125-Luc

Mensch

 

NCI-H1703-Luc-mCh-Puro

Mensch

 

NCI-H1975-Luc

Mensch

 

NCI-H460-Luc2

Mensch

 

PC-9-Luc-mCh-puro

Mensch

Lymphoma

EL4-Luc-mCh-Puro

Maus

Lymphom [B-Cell]

A20-Luc2-Puro

Maus

Lymphom [Burkitt]

Daudi-Luc-mCh-Puro

Mensch

 

Raji-Luc

Mensch

 

Ramos-Luc

Mensch

Lymphom [Diffus gemischt]

SU-DHL-6-Luc-mCh-Puro

Mensch

Lymphom [DLBCL]

OCI-Ly19-Luc-Neo

Mensch

 

OCI-Ly3-Luc-mCh-Puro

Mensch

 

OCI-Ly7-Luc-Neo

Mensch

 

SU-DHL-4-Luc-mCh-Puro

Mensch

 

Toledo-Luc-Neo

Mensch

 

OCI-Ly1 R10-Luc-mCh-Puro

Mensch

 

OCI-Ly1 R7-Luc-mCh-Puro

Mensch

Mamma/Brust

MCF7-Luc-mCh-Puro

Mensch

 

MDA-MB-231-Luc-D3H1

Mensch

 

E0771-Luc-mCh-Puro

Maus

 

MDA-MB-231-Luc-D3H2LN

Mensch

 

EMT6-Luc-mCh-Puro

Maus

 

MX-1-Luc

Mensch

 

4T1-Luc2-1A4

Maus

Melanom

OCM-1-Luc-mCh-Puro

Mensch

 

SK-MEL-28-Luc-mCh-Puro

Mensch

 

B16-F10-Luc-G5

Maus

 

B16-F10-Luc2

Maus

 

Cloudman S91-Luc-mCh-Puro

Maus

 

YUMM1,7-Luc-mCh-Puro

Maus

Myelom

JJN-3-Luc-G418R

Mensch

 

MM.1S (pMMP-Luc-Neo)

Mensch

 

NCI-H929-Luc-mCh-Puro

Mensch

 

5TGM1-Luc

Maus

 

J558-Luc-mCh-Puro

Maus

Ovarial

A2780-Luc

Mensch

 

IGROV1-Luc-Mch-Puro

Mensch

 

OVCAR-5-Luc-mCh-Puro

Mensch

 

OVCAR-8-Luc-mCh-Puro

Mensch

 

SK-OV-3-Luc-D3

Mensch

 

ID8-Luc-mCh-Puro

Maus

 

NIH:OVCAR-3-Luc-mCh-Puro

Mensch

Bauchspeicheldrüse

BxPC-3-Luc2

Mensch

 

MIA PaCa-2-Luc

Mensch

 

PANC-1-Luc

Mensch

 

Pan02.TD1-Luc-mCh-Puro

Maus

Prostata

DU 145-Luc

Mensch

 

PC-3-Luc

Mensch

 

PC-3M-Luc-C6

Mensch

Nieren

293-Luc-mCh-Puro

Mensch

 

786-O-Luc-Neo (gerettet)

Mensch


Table 1: Luciferase-labeled cell lines

Contact us to request the full data set or to learn more about our luciferase enabling service and how it can be applied to your preclinical research.  


Verweise

1 Jessamy C. Tiffen, Charles G. Bailey, Cynthia Ng, John E.J. Rasko & Jeff Holst. Luciferase expression and bioluminescence does not affect tumor cell growth in vitro or in vivo. Molecular Cancer 2010; 9: 299.

2 Dan M. Close, Tingting Xu, Gary S. Sayler, Steven Ripp. In vivo bioluminescent imaging (BLI): noninvasive visualization and interrogation of biological processes in living animals. Sensors (Basel) 2011; 11(1): 180–206.Note: Please note that all animal care and use was conducted according to animal welfare regulations in an AAALAC-accredited facility with IACUC protocol review and approval.

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