Expansion of CD34+ human hematopoietic cells from umbilical cord blood using roller bottles
Expansión de células CD34+ de sangre de cordón umbilical en frascos giratorios
H. Andrade-Zaldívar1,3, M. A. Kalixto-Sánchez2, A. P. Barba de la Rosa1 and A. De León-Rodríguez1*
1 Instituto Potosino de Investigación Científica y Tecnológica (IPICyT), División de Biología Molecular, Camino a la Presa San José, 2055 Col. Lomas 4a sección CP 78216. San Luis Potosí, S.L.P., México. *Corresponding author. E-mail: aleonr@ipicyt.edu.mx Tel. +52-444-8342044, Fax +52-444-8342010.
2 Hospital General del ISSSTE, Carlos Diez Gutiérrez 915 Col. Julian Carrillo CP 78340. San Luis Potosí, S.L.P., México.
]]> 3 Centro de Biociencias de la Universidad Autónoma de San Luis Potosí, Km 14.5 Carretera S.L.P.-Matehuala, Ejido Palma de la Cruz, Soledad de Graciano Sánchez, S.L.P. México.
Received June 11, 2013.
Accepted February 24, 2014.
Abstract
Hematopoietic stem cell (HSC) transplantation is limited by the initial CD34+ cell content in cord blood units; the aim of this work was the in vitro expansion of HSC to overcome this issue. Supplemented IMDM roller bottle cultures of CD34+ cells from human umbilical cord blood were established for hematopoietic progenitor expansion. The maximum total colony forming cells (CFC) expansion was achieved after 5 days of culture, being 16.94±2.82 folds in roller bottles and 17.28±4.47 in static cultures. However, the maximum total cell fold expansion was attained after 10 days of culture. It was of 20.31±6.18 for the roller bottles and 26.45±9.89 for the static cultures. The efficacy of the roller bottles system for short-term cultures of CD34+ cells and expanding hematopoietic progenitors in IMDM was demonstrated; encouraging the use of this culture system for other experiments and may be used for future clinical applications.
Keywords: CD34+, expansion, hematopoietic stem cell, bone marrow transplantation, cell culture, roller bottles.
Resumen
]]> El transplante de células madre hematopoyéticas está limitado por la cantidad de células CD34+ presentes en las unidades de sangre de cordón umbilical; el objetivo de este trabajo fue obtener la expansión in vitro de células madre hematopoyéticas. Se establecieron cultivos de células CD34+ de sangre de cordón umbilical en medio IMDM suplementado con citocinas para promover la expansión de progenitores hematopoyéticos. En 5 días de cultivo en frascos giratorios se obtuvo la máxima expansión de unidades formadoras de colonias (UFC) totales de 16.94±2.82 veces, mientras que en cultivo estático fue de 17.28±4.47. La máxima expansión de células totales fue de 20.31±6.18 veces en frasco giratorio y de 26.45±9.89 veces en cultivo estático a los 10 días de cultivo. Se demostró la eficacia del sistema de frascos giratorios con medio IMDM para el cultivo a corto plazo de células enriquecidas CD34+ provenientes de sangre de cordón umbilical y para la expansión de progenitores hematopoyéticos, potencializando el uso de este sistema para otros experimentos y aplicaciones clínicas a futuro.Palabras clave: CD34+, expansión, células madre hematopoyéticas, trasplantes de médula ósea, cultivo celular, frascos giratorios.
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Acknowledgments
This work was partially supported by CONACYT-Fondos Sectoriales Sector Salud Grant 13993. Dr. Antonio De León Rodríguez thanks the Marcos Moshinky Cathedra 2012 and Hera Andrade thanks the Consejo Nacional de Ciencia y Tecnologia (CONACyT) for the PhD scholarship 103403.
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