Cell-based Assays

DP

 

Draconis Pharma offers a battery of cell-based assays to determine the potential compound role and effect on biological processes within the cell, such as cell proliferation and cytokine production and other cellular responses.

The cell-based assays are performed using immortalized cell lines, primary cell lines or human whole blood.

(for further information or specific requirements, please contact us )

 

 

 

Immortalized cell line assays These commercially available cell lines are stimulated with different stimuli (see table below) in presence or absence of the test compound. After that, cell culture supernatants, total RNA, or cell extracts are collected and assayed for inflammatory mediators.

Human cell type Cell line Stimulus Read-out
Human alveolar epithelial A549 LPS Cytokine expression and/or production
Cell proliferation
Human B cells RAJI/RAMOS LPS Cytokine expression and/or production
Cell proliferation
Human colon epithelium HT29 LPS Cytokine expression and/or production
Cell proliferation
Human monocytic-macrophage , cell line U937, THP1 LPS Cytokine expression and/or production
Cell proliferation
Human T cells Jurkat PMA/ionomycin Cytokine expression and/or production
Cell proliferation

Primary cell line assays
Primary cell lines such as T cells, monocytes, eosinophils, etc are isolated from human whole blood and stimulated in the presence or absence of test compounds. Additionally, experiments may also be performed in combined human cell populations (PBMCs). Cell culture supernatants, total RNA, or cell extracts are collected and assayed for inflammatory mediators.

Primary human cell Stimulus Reference compound Read-out
T-cell (CD3+)

anti-CD3/anti-CD28

IL2

PMA/ionomycin

Mixed lymphocyte reaction

CsA

Jak inhibitor

Tacrolimus

Cytokine expression
and/or production
Monocytes (CD14+) LPS Dexamethasone Cytokine expression
and/or production
Peripheral Blood
Mononuclear Cells (PBMC)

LPS

anti-CD3/anti-CD28

PMA/ionomycin

Dexamethasone

CsA

Cytokine expression
and/or production
Monocyte-derived dendritic
cells (MoDC)

Differentiation with IL4/GMCSF; Maturation with LPS

Cytokine expression
and/or production
Eosinophils

Eotaxin

RANTES

IL3

Chemotaxis

Human whole blood assays
Compounds are also tested in more complex biological systems such as human whole blood. At the end of the experiment, cell culture supernatants, total RNA, or cell extracts are collected and assayed for inflammatory mediators.

Human cells Stimulus Read-out
Human whole blood from healthy volunteers

LPS

anti-CD3/anti-CD28

Cytokine expression and/or production
Human whole blood from RA, psoriasis patients

LPS

anti-CD3/anti-CD28

Cytokine expression and/or production

This assay could be also performed using whole blood from other species

 
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