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Novel culture medium using a small-molecule agonist of thrombopoetin receptor
Poster Title: Novel culture medium using a small-molecule agonist of thrombopoetin receptor
Submitted on 20 Mar 2015
Author(s): Hondo M1; Nishino T2; Inamura M1
Affiliations: 1ReproCELL, Inc. & 2Nissan Chemical Industries
This poster was presented at ISSCR 2014
Poster Views: 1,406
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Poster Information
Abstract: Hematopoietic stem cells (HSCs), defined by their capacity to self-renew and differentiate into all blood cell lineages, can be applied for transplantation therapy. Since a large number of HSCs are required for clinical use, improvement of techniques for expansion of HSCs ex vivo is a critical issue. Several cytokines have been used for this purpose. Thrombopoietin (TPO) is an essential cytokine that regulates megakaryocyte production and HSC proliferation via activating signaling through its receptor c- MPL. We have developed a small-molecule agonist (NR-101) of c-MPL and report that human HSCs are expanded efficiently ex vivo with NR-101. Using a new small-molecule agonist NR- 102 which is related to NR-101, we produced a novel culture medium, ReproHSCTM. The cost for culture of human HSC can be reduced by using this small-molecule.
Here we demonstrated that ReproHSCTM efficiently expands human CD34+CD38- primitive hematopoietic cells in culture and thereby enhances repopulating capacity of HSCs in NOD/SCID mice. Human blood cord CD34+ cells were cultured with ReproHSCTM supplemented with only Stem Cell Factor (SCF) for 7 days. The total cell number was increased about 40-fold during culture. CD34+ cells and CD34+CD38- cells were expanded 12- fold and 8.5-fold, respectively. We then transplanted expanded cells with ReproHSCTM supplemented with SCF and flt3 ligand for 14 days into NOD/SCID mice and analyzed the SCID-repopulating CD45+ cells with flow cytometry. The expanded cells established engraftment better than the fresh CD34+ cells did. These results indicate that ReproHSCTM is a novel medium suitable for the expansion of HSCs ex vivo.
Summary: ReproHSC medium for the culture of hematopoietic stem cells. facilitates the culture and expansion of CD34+/CD38- cells that retain their HSC propertiesReferences: (1) Exp Hematol. 2009 Nov;37(11):1364-1377.e4. Ex vivo expansion of human hematopoietic stem cells by a small-molecule agonist of c-MPL. Nishino T, Miyaji K, Ishiwata N, Arai K, Yui M, Asai Y, Nakauchi H, Iwama A.Report abuse »
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