Umbilical Cord Blood Processing Techniques and Their Comparative Advantages: A Review

Eze Evelyn Mgbeoma *

Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.

Christian Serekara Gideon

Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.

*Author to whom correspondence should be addressed.


Background: Umbilical Cord Blood (UCB) has steadily gained prominence in haematopoietic stem cell transplantation (HSCT). Despite UCB advantages, the main disadvantage of UCB in haematopoietic stem cell transplantation (HSCT) is its limited cell dose. Initially, UCB used to be processed and then made to undergo cryopreservation as whole cord blood banking leading to the problem of storing sufficiently large number of cryoprotected UCB units which requires vast amounts of costly storage space in liquid nitrogen. The sole purpose of processing is to concentrates the stem cells and reduce the volume for storage. Different UCB processing methods have been developed. 

Aim: This review is aimed at bringing together the literature on the different processing methods and highlighting the underlying principles of each method, the relative efficiency and advantages of the methods.

Methodology:  The work involved mainly the critical review of all available academic, professional and industry documents on cord blood processing. The relevant information was obtained from textbooks, academic journals, conference proceedings, the internet among others. The major UCB processing methods include Plasma Depletion, Density Gradient Centrifugation (DGC), Hetastarch, PrepaCyte-CB and Sepax. A study of the potential impact of Hetastarch and PrepaCyte processing methods on transplantation outcomes revealed no difference that was significant was observed between patients receiving cells after the processing regimens were compared.

Results: A comparison of the engraftment time of PrepaCyte-CB with five other processing methods revealed a quicker engraftment time for PrepaCyte-CB processed cord blood units compared to other processing methods. PrepaCyte-CB also recovers significantly more viable stem cells than AutoXpress (AXP) and hydroxyethyl starch (HES) processing methods. Other workers demonstrated that Sepax depletion produces higher recovery of cells that are nucleated. The effect initial volume of cord blood had on the recovery of nucleated cells for the different method of processing were also compared. Recovery when using Sepax is reduced as the unit size processed increases. Hetastarch, which is a density gradient, and plasma depletion separation is also affected in like manner, however, processing done using PrepaCyte-CB was not affected by the initial volume of the collected unit. The advantage of Sepax is that it is fully automated and this allows for mass processing of samples, suitable for bigger cord blood banks. For erythrocyte removal, density gradient separation is a better method that is effective. PrepaCyte-CB is the second most efficient method for removing RBC. The result of Total Nucleated Cells (TNC) and Mono Nucleated Cells (MNC) recovery rate of Hespan and Sepax against AXP processing methods shows that both Hespan and Sepax reproducibly recover greater than 95% of the cord blood stem cells in a typical collection and result in a reduced final volume for final storage.

Conclusion: The five most popular processing techniques are Plasma Depletion, Density Gradient, Hetastarch, PrepaCyte-CB and Automated Centrifugal Machine (Sepax). Most methods involve centrifugation, sedimentation and/or filtration for reducing the red cell content, plasma volume, or both. The different UCB processing methods each has its advantages and disadvantages.

Keywords: Umbilical Cord Blood (UCB), haematopoietic stem-cell transplantation, UCB processing methods, total nucleated cells, mono nucleated cells, cord blood banks

How to Cite

Mgbeoma, E. E., & Gideon, C. S. (2023). Umbilical Cord Blood Processing Techniques and Their Comparative Advantages: A Review. International Blood Research & Reviews, 14(4), 85–98.


Kuljeet S, Ankita S, Nitin M. Subsequent study of the expansion of progenitor stem cells isolated using the best method. Cytotherapy. 2009;11(6):768-77.

Chow R, Lin A, Tonai R. Cell recovery comparison between plasma depletion/reduction- and red cell reduction-processing of umbilical cord blood. Cytotherapy. 2011;13(9):1105–1119.

Rubinstein P. Processing and cryopreservation of placental/umbilical cord blood for unrelated bone marrow reconstitution. Proc. Natl. Acad. Sci. 1995;92:10119–22.

Wise Y. Plasma-depleted versus red cell-reduced umbilical cord blood. Cell Transplantation. 2014;23:407-15

Alonso JM, Regan DM, Johnson CE. A simple and reliable procedure for cord blood banking, processing, and freezing: St Louis and Ohio Cord Blood Bank experiences. Cytotherapy. 2001;3:429–33.

Seyed HM, Morteza Z, Saeid A, Mona A, Bahareh A. Umbilical cord blood quality and quantity: Collection up to transplantation. Asian Journal of Transfusion Science. 2019;13(2):79–89.

Ademokun JA, Chapman C, Dunn J. Lander D, Mair K, Proctor SJ, Dickinson AM. Umbilical cord blood collection and separation for haematopoietic progenitor cell banking. Bone Marrow Transplan. 1997;19(10):1023-8.

Kawasaki-Oyama RB. Blood mesenchymal stem cell culture from the umbilical cord with and without ficoll-paque density gradient method. Scientific Library Online. 2008;1:1.

Basford C, Nicolas F, Saba H, Kendal H, Colin M. The cord blood separation league table: A comparison of the major clinical grade harvesting techniques for cord blood stem cells. International Journal of Stem Cells. 2010;3(1):32-45.

Carroll PD, Christensen RD. New and underutilized uses of umbilical cord blood in neonatal care. Maternal Health, Neonatology and Perinatology. 2015; 1:6-18.

Chow R, Nademanee A, Rosenthal J. Analysis of hematopoietic cell transplants using plasma-depleted cord blood products that are not red blood cell reduced. Biology of Blood and Marrow Transplant. 2007;13:1346–57.

Chow R, Tan P, Jaing T. Hematopoietic stem cell transplantation (HSCT) using plasma depleted umbilical cord blood (UCB) that were not red cell depleted. Blood. 2006;108(11):5231.

Robert YKC, Li Q, Chow C. Cord Blood Stem Cell Processing, Banking and Thawing. eBook (PDF): 2017.

Sivakumaran N, Rathnayaka IR, Shabbir R, Wimalsinghe SS, Jayakody JAS, Chandrasekaran M. Umbilical cord blood banking and its therapeutic uses. International Journal of Scientific Research and Innovative Technology. 2018;5(1):160-172.

Kim G, Kwak J, Kim S. High integrity and fidelity of long-term cryopreserved umbilical cord blood for transplantation. J Clin Med. 2021;10(2):293.

Alois N. Isolation of human lymphocytes by sedimentation. basic exercises in immunochemistry. Springer, Berlin Heidelberg; 1979.

Almici C, Carlo-Stella C, Mangoni L. Density separation of umbilical cord blood and recovery of haemopoietic progenitor cells: Implications for cord blood banking. Stem Cells. 1995;13(5):533-40.

Antoniewicz-Papis J, Dzieciątkowska A, Lachert E. Sedimentation as effective method of preliminary isolation of stem cells from cord blood. Reports of Practical Oncology & Radiotherapy. 2001;6(Supplement 1):S19.

Jaatinen T, Laine J. Isolation of mononuclear cells from human cord blood by ficoll-paque density gradient. Curr. Protoc. Stem Cell Biol; 2007.

Papich MG. Saunders handbook of veterinary drugs small and large animal book, fourth edition, Elsevier USA; 2016.

David H, Diane M, Jeffrey M, Donna M. Umbilical cord blood. AABB Technical Manual. 17th ed. USA; 2011.

Madkaikar M, Gupta M. Ghosh K, Swaminathan S, Sonawane L, Mohanty D. Optimizing methods of red cell sedimentation from cord blood to maximize nucleated cell recovery prior to cryopreservation. Br J Biomed Sci. 2007;64(4):157-9.

Souri M, Zarif MN, Rasouli M, Golzadeh K, Hagh MN, Ezzati N, Atarodi K. Comparison of human umbilical cord blood processing with or without hydroxyethyl starch. Transfusion. 2017;57(11):2758-2766.

Mutiea D, Sartika CR, Dirgantara Y. Cytotherapy. 2018;20(5):e7.

CryoCell International. How is PrepaCyte superior [Internet].


[Accessed On: 2023 May 08].

Basford CN, Forraz S, Habibollah K, McGuckin CP. Umbilical cord blood processing using Prepacyte‐CB increases haematopoietic progenitor cell availability over conventional Hetastarch separation. Cell Prolif. 2009;42(6):751–761.

Lioznov M, Dellbrugger C, Sputtek A, Fehse B, Kroger N, Zander AR. Transportation and cryopreservation may impair haematopoietic stem cell function and engraftment of allogeneic PBSCs, but not BM. Bone Marrow Transplant. 2008;42(2):121–8.

Shoular K, Kaestner A, Kurtzberg J. Umbilical cord blood: Automated sepax processing of large volume units. Stem Cells Translational Medicine-Cord Blood Connect: The International Congress for Cord Blood and Perinatal Tissue Research. 2019;8:S1(S32)-S1-(S35).

Mfarrej B, Vicari O, Ouffai S. Sepax-2 cell processing device: a study assessing reproducibility of concentrating thawed hematopoietic progenitor cells. Journal of Translational Medicine. 2022;20:503.

Risso MA, Deffune E, Luzo CM. Comparison of two methods to process umbilical cord blood into packet red cells for transfusion medicine purposes. Stem Cells Translational Medicine. 2019;8(S1):S29.

Badowski M, Harris D. Collection, processing, and banking of umbilical cord blood stem cells for clinical use in transplantation and regenerative medicine. Methods in Molecular Biology. 2012;879(3):279-90.

Nadimpalli S, Buchanan P, Bloomquist J. Biology of blood and marrow transplantation. Journal of the American society for blood and marrow transplantation. 2017;23(3):S174-S175.

Hanash AM, Levy RB. Donor CD4+CD25+ T cells promote engraftment and tolerance following MHC-mismatched hematopoietic cell transplantation. Blood. 2015;105(4):1828-36.

Lapierre V, Pellegrini N, Bardey I. Cord blood volume reduction using an automated system (Sepax) vs. a semi-automated system (Optipress II) and a manual method (hydroxyethyl starch sedimentation) for routine cord blood banking: A comparative study. Cytotherapy. 2007;9:165–9.

Laroche V, McKenna DH, Moroff G, Schierman T, Kadidlo D, McCullough J. Cell loss and recovery in umbilical cord blood processing: A comparison of post-thaw and post-wash samples. Transfusion. 2005;5:1909–16.

Yoo KH, Lee SH, Kim HJ. The impact of post-thaw colony-forming units-granulocyte/macrophage on engraftment following unrelated cord blood transplantation in pediatric recipients. Bone Marrow Transplant. 2007;39:515–21.

De Kreuk AM, Zevenbergen A, van Oostveen JW, Schuurhuis GJ, Huijgens PC, Jonkhoff AR A single-step colony-forming unit assay for unseparated mobilized peripheral blood, cord blood, and bone marrow. J Hematother Stem Cell Res. 2001;10:795–806.

Chow RYK, Li Q, Chow C. Cord Stem Cell Processing, Banking and Thawing. eBook; 2017.

Regidor C, Posada M, Monteagudo M, Umbilical cord blood banking for unrelated transplantation. Exp Hematol. 1999;27(2):380-5.

Dazey B, Duchez P, Letellier C, Vezon G, Ivanovic Z. Cord blood processing by using a standard manual technique and automated closed system "Sepax" (Kit CS-530). Stem Cells Dev. 2005;14(1):6-10.

Takahashi TA, Rebulla P, Armitage S. Multi-laboratory evaluation of procedures for reducing the volume of cord blood: Influence on cell recoveries. Cytotherapy. 2006;8:254–64.

Lapierre V. Cord blood volume reduction using an automated system (Sepax) versus a semi-automated system (Optipress II) and a manual method (hydroxyethyl starch sedimentation) for routine cord blood banking: A comparative study. Cytotherapy. 2007;9:165–9.

Solves P, Planelles D, Mirabet V, Blanquer A, Carbonell-Uberos F. Qualitative and quantitative cell recovery in umbilical cord blood processed by two automated devices in routine cord blood banking: A comparative study Blood Transfus. 2013;11:405–11.