Although used in academicresearch for several decades, 3D culture models have long been regarded expensive, cumbersome and unnecessary in drug development processes.
Technical advances, coupled with recent observations showing that gene expression in 3D is much closer to clinical expression profiles than those seen in 2D, have renewed attention and generated hope in the feasibility of maturing organotypic 3D systems to therapy test platforms with greater power to predict clinical efficacies.
Here we describe a standardized setup for reproducible, easy-handling culture, treatment and routine analysis of multicellular spheroids, the classical 3D culture system resembling many aspects of the pathophysiological situation in human tumor tissue.
We discuss essential conceptual and practical considerations for an adequate establishment and use of spheroid-based drug screening platforms and also provide a list of human carcinoma cell lines, partly on the basis of the NCI-DTP 60-cell line screen, that produce treatable spheroids under identical culture conditions.
In contrast to many other settings with which to achieve similar results, the protocol is particularly useful to be integrated into standardized large-scale drug test routines as it requires a minimum number of defined spheroids and a limited amount of drug.
The estimated time to run the complete screening protocol described herein–including spheroid initiation, drug treatment and determination of the analytical end points (spheroid integrity, and cell survival through the acid phosphatase assay)–is about 170 h. Monitoring of spheroid growth kinetics to determine growth delay and regrowth, respectively, after drug treatment requires long-term culturing >> or =14 d).
KT 5720 |
B9002-.05 |
ApexBio |
50 µg |
EUR 145 |
KT 5720 |
B9002-.1 |
ApexBio |
100 µg |
EUR 232 |
KT 5720 |
B9002-.25 |
ApexBio |
250 µg |
EUR 483 |
KT 5720 |
B9002-.5 |
ApexBio |
500 µg |
EUR 818 |
KT 5823 (PKG kinase inhibitor) |
SIH-458-100UG |
Stressmarq |
100 µg |
EUR 205 |
|
Description: The substance KT 5823 is a pkg kinase inhibitor. It is synthetically produced and has a purity of ?98%. The pure substance is white solid which is soluble in DMSO (20 mg/ml), dimethyl formamide (20 mg/ml) and ethyl acetate (5 mg/ml). |
KT 5823 (PKG kinase inhibitor) |
SIH-458-500UG |
Stressmarq |
500 µg |
EUR 569 |
|
Description: The substance KT 5823 is a pkg kinase inhibitor. It is synthetically produced and has a purity of ?98%. The pure substance is white solid which is soluble in DMSO (20 mg/ml), dimethyl formamide (20 mg/ml) and ethyl acetate (5 mg/ml). |
Plasminogen Receptor (KT) (PLGRKT) Antibody |
20-abx318476 |
Abbexa |
-
EUR 411.00
-
EUR 1845.00
-
EUR 599.00
-
EUR 182.00
-
EUR 300.00
|
- 100 ug
- 1 mg
- 200 ug
- 20 ug
- 50 ug
|
|
EID1 antibody |
70R-17029 |
Fitzgerald |
50 ul |
EUR 435 |
Description: Rabbit polyclonal EID1 antibody |
Rat Chondroitin Sulfate Epitope 3B3(CS3B3 ELISA Kt |
GA-E0451RT-48T |
GenAsia Biotech |
48T |
EUR 317 |
Rat Chondroitin Sulfate Epitope 3B3(CS3B3 ELISA Kt |
GA-E0451RT-96T |
GenAsia Biotech |
96T |
EUR 496 |
Factor XI Antibody Pair |
55R-FAC11-KT |
Fitzgerald |
5 plates |
EUR 745 |
Description: Factor XI antigen Matched Pair antibody Set for ELISA |
Factor IX Antibody Pair |
55R-FAC9-KT |
Fitzgerald |
5 plates |
EUR 760 |
Description: Factor IX antigen Matched Pair antibody Set for ELISA |
Fibrinogen Matched Pair antibody Set |
55R-FIBRINO-KT |
Fitzgerald |
set |
EUR 833 |
Description: Fibrinogen Matched Pair antibody Set for detection of Human Fibrinogen in the research laboratory |
Mouse RANKL ELISA Kit |
55R-MRANKL-KT |
Fitzgerald |
1 kit |
EUR 658 |
Description: ELISA kit for detection of Mouse RANKL in the research laboratory |
VWF Antibody Pair |
55R-VWF-KT |
Fitzgerald |
5 plates |
EUR 775 |
Description: VWF antigen Matched Pair antibody Set for ELISA |