[IQ Biosciences] Purified Human Monocytes

[IQ Biosciences] Purified Human Monocytes

 

 

 

 

 

SKU: IQB-Hu1-M10

Categories: Human, Normal, PBMC

 

 

 

10 million cells / vial

Monocytes

Frozen

Positively Selected

Human

Normal Peripheral Blood

> 90%

Description

Human Monocytes Application Summary

Purified blood human monocytes are a good source of cells to study the role of monocytes, macrophages, and dendritic cells in the immune response. This is because blood monocytes are commonly used to differentiate into macrophages or dendritic cells with the appropriate cytokine environment. Together, these 3 cell types of the innate immune system serve very important functions, including phagocytosis, cytokine production to enhance the immune response, and antigen presentation to T cells.

The negative isolation of monocytes leaves them untouched without any antibody binding to cell surface markers. This leaves all cell surface proteins eligible to be bound to antibodies or other molecules for functional or population characterization studies.

In contrast, positive isolation of monocytes may lead to internalization of the marker that was used to isolate the cells.  In most cases, these markers are only used for identification purposes and may not have any effect on function, but it will depend on the organism and function.  Therefore, cells isolated using this method may also be employed for functional or population characterization studies with the knowledge that the isolation marker may be internalized and not present.

Human Monocyte Enrichment

Collection of samples

Leukapheresis was performed on verified healthy donors at a world-renowned blood center to obtain PBMCs. PBMCs were carefully washed to remove excess buffers according to iQ Biosciences’ standard operating procedures to obtain clean and healthy PBMCs.

Isolation of Human Monocytes

To enrich for monocytes by the negative isolation method, PBMCs were incubated with antibodies against T cells, B cells, NK cells, dendritic cells, granulocytes, and erythrocytes in a test tube, which was subsequently subjected to a magnet. Cells labeled with the antibodies bound to the magnet through the test tube wall, while unlabeled cells, the monocytes, were decanted into a fresh tube to obtain the enriched population.

For the positive selection method, PBMCs were incubated in a test tube with antibodies against a monocyte cell-specific marker and subsequently subjected to a magnet, similar to negative isolation.  However, the cells that were decanted into the fresh test tube were the non-monocytes, which can be used for other purposes, while the labeled monocytes were left bound to the magnet through the test tube wall.  The test tube was then removed from the magnet to release the purified monocytes directly into the test tube.

Contact us for more information about purchasing our magnetically isolated monocytes.

Purity

Following enrichment, a small aliquot of cells was taken to test for post-sort purity by flow cytometry analysis. Purity of monocytes, as defined by CD14 expression, was > 94% (Figure 1).

Figure 1. Purity of monocytes after negative magnetic isolation from PBMCs.  Human PBMCs were incubated with antibodies against T cells, B cells, NK cells, dendritic cells, granulocytes, and erythrocytes and subjected to negative magnetic isolation to obtain untouched monocytes. A small aliquot was taken after selection to evaluate post-sort purity (right). Percent of CD14+ cells in PBMCs is also shown (left).

Cryopreservation and storage

Purified monocytes 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).