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 < KP < 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.
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References
Albertsson PA, Johansson G, Tjerneld F (1990) Separation processes in biotechnology. Marcel Dekker, New York
Andrade PZ, da Silva CL, dos Santos F, Almeida-Porada G, Cabral JMS (2011) Initial CD34(+) cell-enrichment of cord blood determines hematopoietic stem/progenitor cell yield upon ex vivo expansion. J Cell Biochem. 112:1822–1831
Azevedo AM, Rosa PAJ, Ferreira IF, Aires-Barros MR (2009) Chromatography-free recovery of biopharmaceuticals through aqueous two-phase processing. Trends Biotechnol 27:240–247
Cabral JMS (2007) Cell partitioning in aqueous two-phase polymer systems. In: Kumar A, Galaev IY, Mattiasson B (eds) Cell separation—fundamentals, analytical and preparative methods. Springer-Verlag, Berlin, pp 151–171
Chou S, Chu P, Hwang W, Lodish H (2010) Expansion of human cord blood hematopoietic stem cells for transplantation. Stem Cells. 7:427–428
Delgado C, Malik F, Selisko B, Fischer D, Francis GE (1994) Quantitative analysis of polyethylene glycol (PEG) in PEG-modified proteins/cytokines by aqueous two-phase systems. J Biochem Biophys Meth 29:237–250
Kumar A, Kamihira M, Galaev IY, Mattiasson B, Iijima S (2001) Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers. Biotechnol Bioeng 75:570–580
Mayani H, Lansdorp PM (1998) Biology of human umbilical cord blood-derived hematopoietic stem/progenitor cells. Stem Cells. 16:153–165
Ribeiro SC, Monteiro GA, Cabral JMS, Prazeres DMF (2002) Isolation of plasmid DNA from cell lysates by aqueous two-phase systems. Biotechnol Bioeng 78:376–384
Weissman IL, Shizuru JA (2008) The origins of the identification and isolation of hematopoietic stem cells, and their capability to induce donor-specific transplantation tolerance and treat autoimmune diseases. Blood 112:3543–3553
Wiegel D, Meyer C, Arnold K (1994) Partitioning of high-density lipoproteins in charge-sensitive two-phase systems. J Chrom B Biomed Sci Appl. 661:159–164
Acknowledgments
This work was carried out within the frame of the Project StemSelect (contract no.PTDC/EQU-EPR/104663/2008) granted by Fundação para a Ciência e Tecnologia – FCT. António F. Sousa and Pedro Z. Andrade also acknowledge FCT for grants 3386/2010 and SFRH/BD/38719/2007, respectively.
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Sousa, A.F., Andrade, P.Z., Pirzgalska, R.M. et al. A novel method for human hematopoietic stem/progenitor cell isolation from umbilical cord blood based on immunoaffinity aqueous two-phase partitioning. Biotechnol Lett 33, 2373–2377 (2011). https://doi.org/10.1007/s10529-011-0727-0
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DOI: https://doi.org/10.1007/s10529-011-0727-0