New Method for Sorting Endothelial and Neural Progenitors from Human Induced Pluripotent Stem Cells by Sedimentation Field Flow Fractionation.
Human induced pluripotent stem cells (hiPSc) are a very useful solution to create and observe the behavior of specific and usually inaccessible cells, such as human motor neurons. Obtained from a patient biopsy by reprograming dermal fibroblasts (DF), hiPSc present the same properties as embryonic stem cells and can generate any cell type after several weeks of differentiation. Today, there are numerus protocols which aim to control hiPSC differentiation. The principal challenge is to obtain a sufficiently enriched specific cell population to study disease pathophysiology and to provide a good model for further investigation and drug screening. The differentiation process is very costly and time-consuming, because many specific factors and different culture media must be used. In this study, we used Sedimentation Field Flow Fractionation (SdFFF) to prepare enriched populations derived from hiPSc after only 10 days of culture in a classical medium. Based on phenotypic and proteomic characterization, "hyperlayer" elution resulted in a fraction expressing markers of endothelial progenitors while another fraction expressed markers of neural progenitors. The isolation of subpopulations representing various differentiation lineages is of major interest for the production of specialized, cell-enriched fractions and in the preparation of increasingly complex models for the development of new therapeutic tools.
Pierre-Antoine
PA
Faye
Nicolas
N
Vedrenne
Miguel A
MA
De la Cruz-Morcillo
Claire-Cécile
CC
Barrot
Laurence
L
Richard
Sylvie
S
Bourthoumieu
Franck
F
Sturtz
Département de Génétique, CHU Henri-Mondor , F-94000 Créteil, France.|Inserm U955-E10, Université Paris-Est-Créteil, F-94000 Créteil, France.
Benoît
B
Funalot
Anne-Sophie
AS
Lia
Serge
S
Battu
D002454:Cell Differentiation; D002478:Cells, Cultured; D020405:Dermis / Q000166:cytology; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D031521:Fractionation, Field Flow / Q000379:methods*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008840:Microfilament Proteins / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism
D008840:Microfilament Proteins; D009479:Neuropeptides; D009687:Nuclear Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C112302:ectodermal-neural cortex 1 protein