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Phthalates affect the in vitro expansion of human hematopoietic stem cell

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Abstract

Phthalates are esters of phthalic acid used industrially as plastic additives, however, these are not covalently bound to the polymer matrix and therefore can be released to the environment. The aim of this study was to evaluate the effect of four phthalates: dibutyl phthalate (DBP), benzyl butyl phthalate (BBP), diethyl phthalate (DEP) and diethylhexyl phthalate (DEHP) on the in vitro expansion of human hematopoietic cells from umbilical cord blood. For this, 0.5 × 106 cells/mL were exposure to concentrations ranging from 0.1 to 100 μg/mL and the total cell expansion was determined after 14 days of culture in IMDM-cytokines medium. The control cultures attained 1.31 ± 0.21 × 106 cell/mL, whereas the cultures exposed to DBP, BBP and DEHP showed a reduction from 23 to 81%, 17 to 69% and 15 to 93.5%, respectively. DEP did not affect the total cell expansion. The most significant decrease on total cell expansion was observed at 0.1 μg/mL DBP, 100 μg/mL BBP and 10 μg/mL DEHP (p < 0.05). Additionally, the effect of these compounds on the expansion of hematopoietic progenitors was analyzed by clonogenic assays as colony forming units (CFU). The CFU decreased considerably compared with respect to the control cultures. The reduction was 74.6 and 99.1% at 10 and 100 μg/mL DBP respectively, whereas 100 μg/mL BBP and 100 μg/mL DEHP reduced the CFU expansion in 97.1% and 81%, respectively. Cultures exposed to DEP did not show significant differences. The results demonstrate the toxicity of DBP, BBP and DEHP on the human hematopoietic stem cells.

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Acknowledgements

J. Flores, T. Ortiz, A. Gutiérrez, thank the National Council of Science and Technology (CONACYT) for their scholarships 591428, 298411, 297341, respectively. We would like to thank the partial financial support from CONACyT-Fondo Sectorial SALUD 233340 and CONACyT Problemas Nacionales 2017-4601. We thank Victor E. Balderas for his technical support and the comments and Graham M. Tippett for the English revision.

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  1. Ana K. Gutiérrez-García and José M. Flores-Kelly have contributed equally to this work.

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  1. División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, A.C., Camino a la Presa San José 2055, Col. Lomas 4ª Sección, CP 78216, San Luis Potosí, SLP, Mexico

    Ana K. Gutiérrez-García, José M. Flores-Kelly, Tomás Ortiz-Rodríguez & Antonio De León-Rodríguez

  2. Hospital General del ISSSTE, Carlos Diez Gutiérrez 915, Col. Julian Carrillo, CP 78340, San Luis Potosí, SLP, Mexico

    Marco Antonio Kalixto-Sánchez

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Gutiérrez-García, A.K., Flores-Kelly, J.M., Ortiz-Rodríguez, T. et al. Phthalates affect the in vitro expansion of human hematopoietic stem cell. Cytotechnology 71, 553–561 (2019). https://doi.org/10.1007/s10616-019-00300-x

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