Emergence of CD43-Expressing Hematopoietic Progenitors from Human Induced Pluripotent Stem Cells

Abstract

Background: The ex vivo generation of human hematopoietic stem cells (HSCs) with long-term repopulating capacity and multi-lineage differentiation potential represents the holy grail of hematopoiesis research. In principle, human induced pluripotent stem cells (hiPSCs) provide the tool for both studying molecular mechanisms of hematopoietic development and the ex vivo production of ‘true' HSCs for transplantation purposes and lineage-specific cells, e.g. red blood cells, for transfusion purposes. CD43-expressing cells have been reported as the first hematopoietic cells during development, but whether or not these possess multilineage differentiation and long-term engraftment potential is incompletely understood. Methods: We performed ex vivo generation of hematopoietic cells from hiPSCs using an embryoid body(EB)-based, xeno-product-free differentiation protocol. We investigated the multilineage differentiation potential of different FACS-sorted CD43-expressing cell subsets by colony-forming assays in semisolid media. Further, erythroid differentiation was investigated in more detail using established protocols. Results: By using CD43, we were able to measure hematopoietic induction efficiency during hiPSC-derived EB differentiation. Further, we determined CD43+ cells as the cell population of origin for in vitro erythropoiesis. Furthermore, colony formation demonstrates that the multipotent hematopoietic stem and progenitor cell fraction is particulary enriched in the CD43^hi CD45+ population.

Multilineage differentiation potential of hematoendothelial progenitors derived from human induced pluripotent stem cells

“…The day 8 HSPCs were efficiently differentiated to erythroid cells expressing γ-, ε-, and low levels of β- globin with a small proportion of the cells enucleated and became reticulocytes. The obtained erythroid cells were similar to those from previous studies, which does not represent adult red cell phenotype [46][47][48]. In this study, we detected the expression of various transcription factors known to be related to erythropoiesis during erythroid differentiation; however, BCL11A was not expressed throughout the differentiation.…”

Decoding human fetal liver haematopoiesis

“…We explored supporting factors for fetal liver erythropoiesis using CellPhoneDB 16 to predict specific/enriched receptor-ligand interactions between erythroblasts and VCAM1 + EI macrophages (Extended Data 4a). We identify statistically significant interactions for VCAM1, ITGB1, ITGA4, SIGLEC1, ICAM4 and SPN, molecules known to be important in haematopoiesis (Extended Data 4a) 17,18 . The presence of VCAM1 on EI macrophages and ITGA4 on early/mid erythroid cells is confirmed by immunohistochemical analysis on serial fetal liver sections (Extended Data 4b).…”

“…After 8 PCW, developmental age was estimated from measurements of foot length and heel to knee length and compared against a standard growth chart 53 . A piece of skin, or where this was not possible, chorionic villi tissue was collected from every sample for Quantitative Fluorescence-Polymerase Chain Reaction analysis using markers for the sex chromosomes and the following autosomes 13,15,16,18,21,22, which are the most commonly seen chromosomal abnormalities. All samples were karyotypically normal.…”

Induced Pluripotent Stem Cell-Derived Red Blood Cells and Platelet Concentrates: From Bench to Bedside

“…The differentiated cells in most reports expressed fetal and embryonic globins, indicating reprogramming of the globin locus from the original parental cell. Erythroid differentiation has been confirmed by morphological analysis and expression of a very limited number of RBCs markers, including sialophorin (CD43) [ 37 ], glycophorin A, and transferrin receptor [ 34 , 35 ]. The reticulocytes generated from iPS cells exhibit an oxygen binding capacity similar to cord blood RBCs, which contain predominantly fetal hemoglobin [ 36 ], however, the dynamics of the gene expression program (largely erased during reprogramming) found during erythropoiesis from hiPS cells was found distinct compared with adult and cord blood progenitors [ 38 ].…”