ASSAY – DC MATURATION
HUMAN CELL LINE BASED
MONOCYTE DERIVED DENDRITIC CELLS
Dendritic cells (DCs) are the star antigen presenting cells (APCs) of myeloid or haematopoietic origin that play a key role between the innate and adaptive immune systems. DCs are pivotal in the T cell mediated T-helper response to MHC ClassII-Ag complex, and also cross presenting via the MHC ClassI pathway. They consist of receptors for pathogen and danger associated molecular patterns e.g. TLRs.
In vitro Dendritic Cell (DC) Maturation Studies
We generate DCs from a human monocytic cell line (also from primary monocytes or haematopoietic cells). These cells express CD11c, HLA-DR, CD80, CD86, SIRPa, CD14 and can be used in mono or co-cultures to study drug effects for DC maturation and various DC related outcomes.
The DCs can be characterised by flow cytometry, immunohistochemistry and microscopy and also through cytokine measurements. We can perform assays with a T cell line for immune functional studies.
DCs are very important in studies of various therapeutic areas and biological pathways:
- Tumour biology and oncology
- Wound healing
- Inflammatory and autoimmune conditions
- Cardiovascular disease
- Tissue damage and cell death
In recent years much attention has been given to in vitro DC based models in cancer studies.
Cell Line Based Approach
We use myeloid and lymphocytic cell lines for various immune response assays such as dendritic cell maturation, T cell activation, phagocytosis, etc. We are among the few groups who can generate and demonstrate the use of dendritic cells and macrophages (including M1 and M2) from monocytic cell lines and T cells in activation studies. Conventionally dendritic cells are generated from primary monocytes or bone marrow and T cells or monocytes are separated from primary PBMCs from human donors. The use of cell lines in the initial stage offers a consistent reference assay prior to using primary human cells. Primary human cells used in the initial stage can be variable, expensive, time consuming, and often inconclusive.