Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
Por:
Mora-Roldan G.A., Ramirez-Ramirez, Dalia, Pelayo, Rosana, Gazarian, Karlen
Publicada:
1 nov 2021
Categoría:
Biochemistry, genetics and molecular biology (miscellaneous)
Resumen:
Background. In vitro methods for hematopoietic differentiation of human
pluripotent stem cells (hPSC) are a matter of priority for the in-depth
research into the mechanisms of early embryogenesis. So-far, published
results regarding the generation of hematopoietic cells come from
studies using either 2D or 3D culture formats, hence, it is difficult to
discern their particular contribution to the development of the concept
of a unique in vitro model in close resemblance to in vivo
hematopoiesis. Aim of the study. To assess using the same culture
conditions and the same time course, the potential of each of these two
formats to support differentiation of human pluripotent stem cells to
primitive hematopoiesis without exogenous activation of Wnt signaling.
Methods. We used in parallel 2D and 3D formats, the same culture
environment and assay methods (flow cytometry, IF, qPCR) to investigate
stages of commitment and specification of mesodermal, and hemogenic
endothelial cells to CD34 hematopoietic cells and evaluated their
clonogenic capacity in a CFU system. Results. We show an adequate
formation of mesoderm, an efficient commitment to hemogenic endothelium,
a higher number of CD34 hematopoietic cells, and colony-forming capacity
potential only in the 3D format-supported differentiation. Conclusions.
This study shows that the 3D but not the 2D format ensures the induction
and realization by endogenous mechanisms of human pluripotent stem
cells' intrinsic differentiation program to primitive hematopoietic
cells. We propose that the 3D format provides an adequate level of
upregulation of the endogenous Wnt/beta-catenin signaling.
Filiaciones:
Mora-Roldan G.A.:
Departamento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Cuidad de México, 04510, Mexico
Instituto de Oftalmología Fundación, Conde de Valenciana IAP, Cuidad de Mexico, 04510, Mexico
Ramirez-Ramirez, Dalia:
Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Km 4.5 Carretera Atlixco-Metepec, Puebla, 74360, Mexico
Inst Mexicano Seguro Social, Ctr Invest Biomed Oriente, Km 4-5 Carretera Atlixco Metepec, Puebla 74360, Mexico
Pelayo, Rosana:
Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Km 4.5 Carretera Atlixco-Metepec, Puebla, 74360, Mexico
Inst Mexicano Seguro Social, Ctr Invest Biomed Oriente, Km 4-5 Carretera Atlixco Metepec, Puebla 74360, Mexico
Gazarian, Karlen:
Departamento de Medicina Genómica y Toxicología Ambiental, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Cuidad de México, 04510, Mexico
Univ Nacl Autonoma Mexico, Dept Med Genom & Toxicol Ambiental, Inst Invest Biomed, Cuidad De Mexico 04510, Mexico
Inst Oftalmol Fdn, Conde Valenciana IAP, Cuidad De Mexico 04510, Mexico
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