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Updated: Aug 13, 2026
Generation of Human Monocyte-derived Dendritic Cells from Whole Blood
Published on: December 24, 2016
Storage of blood for in vitro generation of dendritic cells
R Syme1, D Callaghan, P Duggan
1Department of Oncology, Faculty of Medicine, University of Calgary, Tom Baker Cancer Center, Calgary, Canada.
Insights
Storing blood at room temperature for 24 hours preserves dendritic cell (DC) yield and morphology for immunotherapies. Refrigeration reduces cell numbers and affects DC phenotype, making room temperature storage optimal when immediate processing isn't feasible.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are crucial for immunotherapy.
- Clinical applications require processing blood after collection.
- Delays in blood processing can impact DC quality.
Purpose of the Study:
- To evaluate the impact of overnight blood storage on DC yield, morphology, and phenotype.
- To determine optimal storage conditions for maintaining DC viability and function.
Main Methods:
- Peripheral blood mononuclear cells (PBMCs) were processed immediately or after 24-hour storage at 4°C (refrigerated) or 20°C (room temperature).
- Assessed starting cell number, DC yield, morphology, and phenotype (MHC II expression).
Main Results:
- Refrigeration significantly reduced PBMC numbers compared to fresh or room temperature storage.
- Room temperature storage yielded DCs with morphology identical to fresh samples.
- Refrigerated samples showed reduced MHC II expression, impacting DC phenotype.
- Total DC yield and percentage yield were not significantly affected by storage conditions.
Conclusions:
- Overnight storage at room temperature (20°C) is recommended if immediate blood processing for in vitro DC generation is not possible.
- Refrigeration (4°C) negatively impacts PBMC yield and DC phenotype (MHC II expression).
Background:
Since the development of techniques to cultivate DC from peripheral blood, there has been a great deal of interest in the use of these cells in immunotherapeutic strategies. In a clinical setting, delays often occur between when blood is drawn and when it is processed. We therefore investigated the effect of overnight storage on the yield, morphology and phenotype of DC cultured from the peripheral blood of healthy volunteers.
Method:
Blood was processed either immediately, or after storage for 24 h in the fridge (4 degrees C) or at room temperature (RT, 20 degrees C). Samples were compared for starting cell number, DC yield and characteristics (morphology and phenotype).
Results:
The number of PBMC that could be obtained was significantly lower from the refrigerated samples compared with both the freshly processed sample and that stored at RT. Samples processed after overnight storage at RT yielded cells morphologically identical to DC cultured from freshly processed samples. Only when samples were both stored and processed cold did the cultured cells not have typical DC morphology. DC cultured from the refrigerated samples showed a significant reduction in MHC II expression compared with samples processed fresh or stored at RT. This expression increased slightly when the sample was first warmed. Total DC yield and the percentage yield of cultured DC was not significantly different for any of the groups.
Discussion:
We conclude that, if immediate processing of blood for in vitro generation of DC is not possible, samples should be stored at room temperature (approximately 20 degrees C).
