Efficient Expansion of SALL4–Transduced Umbilical Cord Blood Derived CD133+Hematopoietic Stem Cells
Hematopoietic stem cells (HSCs) were characterized by self-renewal and multilineage potential. Umbilical cord blood-derived (UCB) as an alternative source of HSCs is widely used especially in children for stem cells transplant (SCT). The main limitation in using UCB for transplantation especially in adults is low cell dose. To overcome this limitation besides using double dose UCB, ex vivo expansion is the most important way to increase cell number for transplantation. HSCs are mainly isolated using CD133 or CD34. CD133, as the most primitive marker, shows important physiological role in maintenance and expansion of HSCs. SALL4 plays crucial role in the development and maintaining the pluripotency and self-renewal ability of embryonic stem cells (ESCs) as well as HSCs. Moreover, SALL4 act as a regulator of HSCs expansion, normal hematopoiesis, and hematological malignancies. In the present study, CD133+ cells positively selected and ex vivo expanded in SALL-4 and GFP-transduced group. CD133 expression assessed using flow cytometry at day 0, 7 and 10. Moreover, multilineage differentiation and proliferation potential of expanded cells in both groups evaluated using colony forming unit (CFU) assay, and cells count assay. Karyotyping analysis was performed to assess any chromosomal instability after 7 days of expansion. Obtained results demonstrated that SALL-4 transduced cells showed significant increase in cell number compared to control group. Moreover, immunophenotyping results showed higher expression level of CD133 at day 7 and 10 following expansion in SALL-4 transduced (62 % and 42%) compared to control group (51% and 20.6%). Our results illustrated that SALL4 could act as a positive factor for the expansion of CD133+ derived UCB cells besides maintaining self-renewal and differentiation ability of expanded cell without any numerical and structural chromosomal aberrations.
Gluckman E, Broxmeyer HA, Auerbach AD, Friedman HS, Douglas GW, Devergie A, et al. Hematopoietic reconstitution in a patient with Fanconi's anemia by means of umbilical-cord blood from an HLA-identical sibling. The New England journal of medicine. 1989;321(17):1174-8. Epub 1989/10/26.
Mayani H. Umbilical cord blood: lessons learned and lingering challenges after more than 20 years of basic and clinical research. Archives of medical research. 2011;42(8):645-51. Epub 2011/12/14.
Flores-Guzman P, Fernandez-Sanchez V, Mayani H. Concise review: ex vivo expansion of cord blood-derived hematopoietic stem and progenitor cells: basic principles, experimental approaches, and impact in regenerative medicine. Stem cells translational medicine. 2013;2(11):830-8. Epub 2013/10/09.
Schuster JA, Stupnikov MR, Ma G, Liao W, Lai R, Ma Y, et al. Expansion of hematopoietic stem cells for transplantation: current perspectives. Experimental hematology & oncology. 2012;1(1):12. Epub 2012/12/06.
Ren Z, Jiang Y. Umbilical cord blood hematopoietic stem cell expansion ex vivo. Journal of Blood Disorders & Transfusion. 2013;2012.
Tung SS, Parmar S, Robinson SN, De Lima M, Shpall EJ. Ex vivo expansion of umbilical cord blood for transplantation. Best practice & research Clinical haematology. 2010;23(2):245-57. Epub 2010/09/15.
Andrade-Zaldivar H, Santos L, De Leon Rodriguez A. Expansion of human hematopoietic stem cells for transplantation: trends and perspectives. Cytotechnology. 2008;56(3):151-60. Epub 2008/11/13.
Mizrak D, Brittan M, Alison M. CD133: molecule of the moment. The Journal of pathology. 2008;214(1):3-9. Epub 2007/12/11.
Antonchuk J, Sauvageau G, Humphries RK. HOXB4-induced expansion of adult hematopoietic stem cells ex vivo. Cell. 2002;109(1):39-45. Epub 2002/04/17.
Aggarwal R, Pompili VJ, Das H. Genetic modification of ex-vivo expanded stem cells for clinical application. Front Biosci (Landmark Ed). 2010;15:854-71. Epub 2010/06/03.
Aguila JR, Liao W, Yang J, Avila C, Hagag N, Senzel L, et al. SALL4 is a robust stimulator for the expansion of hematopoietic stem cells. Blood. 2011;118(3):576-85. Epub 2011/05/24.
Kohlhase J, Schuh R, Dowe G, Kuhnlein RP, Jackle H, Schroeder B, et al. Isolation, characterization, and organ-specific expression of two novel human zinc finger genes related to the Drosophila gene spalt. Genomics. 1996;38(3):291-8. Epub 1996/12/15.
Akhavan Rahnama M, Movassaghpour AA, Soleimani M, Atashi A, Anbarlou A, Shams Asenjan K. MicroRNA-15b target Sall4 and diminish in vitro UCB-derived HSCs expansion. EXCLI journal. 2015;14:601-10. Epub 2015/12/10.
Yang J, Liao W, Ma Y. Role of SALL4 in hematopoiesis. Current opinion in hematology. 2012;19(4):287-91. Epub 2012/05/05.
Yang JC, Chai L, Gao C, Fowles TC, Alipio Z, Dang HE, et al. SALL4 is a key regulator of survival and apoptosis in human leukemic cells. Blood. 2008;112(3):805-13.
Zhang X, Yuan X, Zhu W, Qian H, Xu WR. SALL4: An emerging cancer biomarker and target. Cancer Lett. 2015;357(1):55-62.
Aguila JR, Mynarcik DC, Ma Y. SALL4: finally an answer to the problem of expansion of hematopoietic stem cells? Expert review of hematology. 2011;4(5):479-81. Epub 2011/09/24.
Katayama Y, Miyamoto K, Takenaka K, Imajyo K, Shinagawa K, Harada M. Chromosome analysis after ex vivo expansion of CD34(+) cells from human cord blood. Cancer genetics and cytogenetics. 2001;125(2):161-2. Epub 2001/05/23.
Robinson SN, Ng J, Niu T, Yang H, McMannis JD, Karandish S, et al. Superior ex vivo cord blood expansion following co-culture with bone marrow-derived mesenchymal stem cells. Bone marrow transplantation. 2006;37(4):359-66. Epub 2006/01/10.
Koutna I, Peterkova M, Simara P, Stejskal S, Tesarova L, Kozubek M. Proliferation and differentiation potential of CD133+ and CD34+ populations from the bone marrow and mobilized peripheral blood. Annals of hematology. 2011;90(2):127-37. Epub 2010/09/08.
Brendel C, Goebel B, Daniela A, Brugman M, Kneissl S, Schwable J, et al. CD133-targeted gene transfer into long-term repopulating hematopoietic stem cells. Molecular therapy : the journal of the American Society of Gene Therapy. 2015;23(1):63-70. Epub 2014/09/06.
Bari S, Seah KK, Poon Z, Cheung AM, Fan X, Ong SY, et al. Expansion and homing of umbilical cord blood hematopoietic stem and progenitor cells for clinical transplantation. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation. 2015;21(6):1008-19. Epub 2015/01/04.
Shen B, Zhang Y, Dai W, Ma Y, Jiang Y. Ex-vivo expansion of nonhuman primate CD34+ cells by stem cell factor Sall4B. Stem cell research & therapy. 2016;7(1):152. Epub 2016/10/22.
Milanovich S, Peterson J, Allred J, Stelloh C, Rajasekaran K, Fisher J, et al. Sall4 overexpression blocks murine hematopoiesis in a dose-dependent manner. Experimental hematology. 2015;43(1):53-64 e1-8. Epub 2014/09/24.
Ge J, Cai H, Tan WS. Chromosomal stability during ex vivo expansion of UCB CD34(+) cells. Cell proliferation. 2011;44(6):550-7. Epub 2011/10/07.
Tian H, Huang S, Gong F, Tian L, Chen Z. Karyotyping, immunophenotyping, and apoptosis analyses on human hematopoietic precursor cells derived from umbilical cord blood following long-term ex vivo expansion. Cancer genetics and cytogenetics. 2005;157(1):33-6. Epub 2005/01/29.
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