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Updated: May 30, 2026
Isolation of Precursor B-cell Subsets from Umbilical Cord Blood
Published on: April 16, 2013
A novel method for human hematopoietic stem/progenitor cell isolation from umbilical cord blood based on
António F Sousa1, Pedro Z Andrade, Roksana M Pirzgalska
1Department of Bioengineering, Institute for Biotechnology and Bioengineering, Centre for Biological and Chemical Engineering, Technical University of Lisbon, Av. Rovisco Pais, 1049-001 Lisbon, Portugal.
Insights
This study presents a new method using immunoaffinity aqueous two-phase systems to isolate CD34(+) stem cells from umbilical cord blood. The process efficiently enriches these vital cells for potential clinical applications.
Area of Science:
- Biotechnology
- Cell Biology
- Hematology
Background:
- Hematopoietic stem/progenitor cells (HSPCs) are crucial for treating various diseases.
- Current methods for isolating CD34(+) cells from umbilical cord blood (UCB) can be complex and may reduce cell viability.
- Efficient and direct isolation of CD34(+) cells from UCB is needed for clinical applications.
Purpose of the Study:
- To develop and evaluate a novel immunoaffinity aqueous two-phase system (ATPS) for direct isolation and purification of CD34(+) stem/progenitor cells from whole UCB.
- To assess the efficiency of selective CD34(+) cell recovery and purification using this ATPS.
Main Methods:
- Utilized an ATPS composed of polyethylene glycol (PEG) and dextran for cell separation.
- Employed a monoclonal antibody targeting the CD34 surface antigen to facilitate selective partitioning of CD34(+) cells.
- Analyzed the enrichment of CD34(+) cells and partitioning of contaminant cells in the PEG-rich and dextran-rich phases, respectively.
Main Results:
- Achieved a significant enrichment of CD34(+) cells from an initial 0.2% to up to 42% in a single partitioning step.
- Demonstrated high recovery yield of 95% for CD34(+) cells.
- Reported a high purification factor of 245 for CD34(+) cells, with contaminant cells partitioning to the dextran-rich phase.
Conclusions:
- The immunoaffinity ATPS offers a novel, efficient, and direct method for purifying CD34(+) stem/progenitor cells from whole UCB.
- This method shows potential for improving HSPC isolation for various clinical settings.
- The high recovery and purification achieved suggest a promising alternative to existing cell separation techniques.
Abstract:
A novel cell separation process based on immunoaffinity aqueous two phase systems is presented to isolate and purify CD34(+) stem/progenitor cells directly from the whole umbilical cord blood (UCB). A system, composed of polyethylene glycol and dextran, was evaluated for the selective recovery of CD34(+) cells from UCB. A monoclonal antibody against the CD34 surface antigen was used for the direct partitioning of CD34(+) cells in UCB to the PEG-rich phase. The initial population of CD34(+) cells (0.2% of the initial sample) was enriched to values up to 42% in a single partitioning step, while the majority of contaminant cells were partitioned to the dextran-rich phase (1.37 × 10(-2) < K(P) < 2.76 × 10(-2)). This novel selection method allowed a recovery yield of 95% of CD34(+) cells with a purification factor of 245 and is expected to pave a new way to purify hematopoietic stem/progenitor cells for use in a variety of clinical settings.
