Description

About the Rhesus Monkey (or Rhesus Macaque)

Rhesus macaques are one of the most commonly utilized non-human primates in biomedical research. They are employed in numerous research areas, such as immunology, neuroscience, oncology, infectious disease, and toxicology due to their physiology.

Rhesus Macaque Monkey CD14+ Cells

CD14+ cells make up approximately a tenth of cells in the blood of a typical adult rhesus monkey, and monocytes and neutrophils typically make up the cells that express CD14+ cells. When activated, monocytes can phagocytose foreign cells and perform antibody-dependent cellular phagocytosis (ADCP), as well as produce and secrete inflammatory cytokines.

Developmentally, monocytes can mature into macrophages and dendritic cells depending on the soluble factors they are exposed to in order to become more specialized immune subsets.

Finally, neutrophils are a subtype of granulocytes, which can also perform phagocytosis of microorganisms and secrete enzymes to destroy them.

Rhesus CD14+ Cell Application Summary

Purified rhesus CD14+ cells are a good source of cells to study the biology of monocytes and their role in the immune system since they make up approximately 50% of CD14+ cells. These isolated cells can be used to examine the differentiation and development of the monocyte population to the macrophage or dendritic cell subsets depending on the cytokine environment. Further, these differentiated cells and monocytes can also be used for stimulation-dependent functional experiments to assess cytokine production and antigen presentation.

Similarly, isolated CD14+ cells are a good source of mature neutrophils to study their biology, in particular their functional capabilities. They are capable of secreting enzymes, as well as performing phagocytosis in response to foreign organisms.

With the current trend toward development of immuno-modulatory drugs, CD14+ cells are an ideal source of cells to test inflammatory factor production in the presence of therapeutic molecules. More importantly, these cells can be used to assess toxicity and safety characteristics of these biologics before first in-human trials as these cells can be employed in pre-clinical settings to ensure the biologics are not eliciting unwanted monocyte or neutrophil-mediated functions.

Rhesus CD14+ Cell Purification

Collection of samples

Rhesus PBMCs were sourced from a responsible third-party vendor that operates according to local regulations and laws.

Magnetic Positive Isolation of CD14+ Cells

To purify rhesus CD14+ cells, PBMCs were incubated with antibodies against CD14 and subsequently passed through a magnetic column. The cells that pass through are the non-CD14+ cells, which can be used for other purposes, while the labeled CD14+ cells are left bound to the column. These cells are subsequently eluted to obtain purified CD14+ cells.

Typically, CD14+ make up 10-15% of the total rhesus PBMC populations (Figure 1, left). After isolating through the magnetic columns, the purity is >95% (Figure 1, right).

Figure 1. Flow profiles of rhesus CD14+ cells pre and post purification.  Briefly, rhesus PBMCs were incubated with CD14+ microbeads and passed through a magnetic column.  The magnetic-bound cells were then eluted after a number of washes to remove contaminating cells.  Flow profiles of CD14 staining pre- (left) and post-purification (right) are shown.  CD14+ percentage is listed on the top right of the flow profile.

Cryopreservation and storage

Purified rhesus CD14+ cells were cryopreserved carefully using iQ Biosciences’ cryopreservation protocol that ensures high viability (> 90%) after thawing.

Cells should be stored at < -120 °C once they are received, such as within a liquid nitrogen tank (vapor phase).

Contact us for more information about purchasing our magnetically isolated rhesus CD14+ cells.