ルシフェラーゼ標識化細胞株:オンコロジー研究の貴重なツール

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.

組織型

Cell line

D54-Luc

ヒト

 

Gli36-DsRed-R-Luc(レスキュー)

ヒト

 

LN-827(pMMP-LucNeo)

ヒト

 

U-251-Luc-mCh-Puro

ヒト

 

U-87 MG-Luc

ヒト

 

GL261-Luc2

マウス

 

9L-Luc

ラット

膀胱

T24-Luc-Neo

ヒト

 

MB49-Luc-mCh-Puro

マウス

結腸

COLO 205-Luc #2

ヒト

 

HCT-116-Luc

ヒト

 

HT-29-Luc

ヒト

 

CT26.WT-luc-mCh-puro

マウス

 

MC38-NCI.TD1-luc-mCh-puro

マウス

Endometrial

KLE-Luc-mCh-Puro

ヒト

白血病 [AML]

Kasumi-3-Luc-mCh-Puro

ヒト

 

KG-1-Luc-mCh-Puro

ヒト

 

MV-4-11-Luc-mCh-Puro

ヒト

 

C1498-Luc-mCh-Puro

マウス

白血病 [B-ALL]

NALM6-Luc-MCh-Puro

ヒト

 

Reh (pMMP-Luc-Neo)

ヒト

白血病 [CML]

K-562-Luc2

ヒト

Leukemia [erythro]

HEL 92.1.7-Luc-Neo

ヒト

 

HEL-Luc-Neo

ヒト

白血病 [T-ALL]

DND-41-Luc-mCh-Puro

ヒト

 

MOLT-4-Luc-MCh-Puro

ヒト

肝臓

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

マウス

 

Hep-55.1C-Luc-mCh-Puro

マウス

 

Hepa 1-6-Luc-mCh-Puro

マウス

LL/2-Luc-M38

マウス

 

AB1-Luc-mCh-puro

マウス

肺 [NSCLC]

A549-Luc-C8

ヒト

 

HCC827-Luc-mCh-Puro

ヒト

 

NCI-H125-Luc

ヒト

 

NCI-H1703-Luc-mCh-Puro

ヒト

 

NCI-H1975-Luc

ヒト

 

NCI-H460-Luc2

ヒト

 

PC-9-Luc-mCh-puro

ヒト

リンパ腫

EL4-Luc-mCh-Puro

マウス

リンパ腫 [B 細胞]

A20-Luc2-Puro

マウス

リンパ腫 [バーキット]

Daudi-Luc-mCh-Puro

ヒト

 

Raji-Luc

ヒト

 

Ramos-Luc

ヒト

リンパ腫 [びまん性混合型]

SU-DHL-6-Luc-mCh-Puro

ヒト

リンパ腫 [DLBCL]

OCI-Ly19-Luc-Neo

ヒト

 

OCI-Ly3-Luc-mCh-Puro

ヒト

 

OCI-Ly7-Luc-Neo

ヒト

 

SU-DHL-4-Luc-mCh-Puro

ヒト

 

Toledo-Luc-Neo

ヒト

 

OCI-Ly1 R10-Luc-mCh-Puro

ヒト

 

OCI-Ly1 R7-Luc-mCh-Puro

ヒト

乳房/乳腺

MCF7-Luc-mCh-Puro

ヒト

 

MDA-MB-231-Luc-D3H1

ヒト

 

E0771-Luc-mCh-Puro

マウス

 

MDA-MB-231-Luc-D3H2LN

ヒト

 

EMT6-Luc-mCh-Puro

マウス

 

MX-1-Luc

ヒト

 

4T1-Luc2-1A4

マウス

黒色腫

OCM-1-Luc-mCh-Puro

ヒト

 

SK-MEL-28-Luc-mCh-Puro

ヒト

 

B16-F10-Luc-G5

マウス

 

B16-F10-Luc2

マウス

 

Cloudman S91-Luc-mCh-Puro

マウス

 

YUMM1.7-Luc-mCh-Puro

マウス

骨髄腫

JJN-3-Luc-G418R

ヒト

 

MM.1S (pMMP-Luc-Neo)

ヒト

 

NCI-H929-Luc-mCh-Puro

ヒト

 

5TGM1-Luc

マウス

 

J558-Luc-mCh-Puro

マウス

卵巣

A2780-Luc

ヒト

 

IGROV1-Luc-Mch-Puro

ヒト

 

OVCAR-5-Luc-mCh-Puro

ヒト

 

OVCAR-8-Luc-mCh-Puro

ヒト

 

SK-OV-3-Luc-D3

ヒト

 

ID8-Luc-mCh-Puro

マウス

 

NIH:OVCAR-3-Luc-mCh-Puro

ヒト

膵臓

BxPC-3-Luc2

ヒト

 

MIA PaCa-2-Luc

ヒト

 

PANC-1-Luc

ヒト

 

Pan02.TD1-Luc-mCh-Puro

マウス

前立腺

DU 145-Luc

ヒト

 

PC-3-Luc

ヒト

 

PC-3M-Luc-C6

ヒト

腎臓

293-Luc-mCh-Puro

ヒト

 

786-O-Luc-Neo(レスキュー)

ヒト


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.  


参照

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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