- Human monocytes are purified from PBMCs of healthy blood donors sourced from a world-renowned blood center
- Greater than 90% post-selection purity achieved via magnetic isolation
- Carefully cryopreserved to ensure high viability (> 90%) upon thawing
- All orders come with an iQ Certificate of Analysis
Purified Human Monocytes
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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.
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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).
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We are Committed to Ethical Practices
iQ Biosciences’ human primary cell products are lawfully obtained in accordance with Local, State, and Federal U.S. requirements, and the collection of cells complies with ethical requirements. Our cells are obtained from normal or disease patient volunteers participating in a donor program that is approved by an Institutional Review Board (IRB) or Human Subject Committee. A signed and witnessed consent form is obtained from donor volunteers prior to starting the collection protocol. Strict controls on personal identifiers of volunteers are in place in order to protect their privacy.
For US customers, we ship via FedEx Overnight Shipping. Shipping charges will vary per shipping address (based on ZIP code) and are estimated to be $110.
For international (non-US) customers, we work closely with you and our couriers to ensure all necessary documentation is in place for international shipments to significantly reduce the chance of delays at Customs. For the export of non-human primate samples, this includes preparing CITES permits, as well as any other documentation as required by country. Please submit an inquiry to firstname.lastname@example.org for your estimated time of delivery and shipping charges.
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