Production of Embryonic and Fetal-Like Red Blood Cells from Human Induced Pluripotent Stem Cells

Abstract

We have previously shown that human embryonic stem cells can be differentiated into embryonic and fetal type of red blood cells that sequentially express three types of hemoglobins recapitulating early human erythropoiesis. We report here that we have produced iPS from three somatic cell types: adult skin fibroblasts as well as embryonic and fetal mesenchymal stem cells. We show that regardless of the age of the donor cells, the iPS produced are fully reprogrammed into a pluripotent state that is undistinguishable from that of hESCs by low and high-throughput expression and detailed analysis of globin expression patterns by HPLC. This suggests that reprogramming with the four original Yamanaka pluripotency factors leads to complete erasure of all functionally important epigenetic marks associated with erythroid differentiation regardless of the age or the tissue type of the donor cells, at least as detected in these assays. The ability to produce large number of erythroid cells with embryonic and fetal-like characteristics is likely to have many translational applications.

Altered hematopoiesis in trisomy 21 as revealed through in vitro differentiation of isogenic human pluripotent cells

“…However, during routine quality-control analysis of serially passaged trisomic DS1-iPS4 iPS cells, we observed the emergence of a mixed population consisting of cells disomic and trisomic for chromosome 21. DNA microsatellite analysis excluded inadvertent contamination of the original trisomic line.…”

“…TMD and AMKL in trisomy 21 individuals represent disorders of fetal hematopoiesis. Prior experience with human pluripotent cells has revealed a strong bias toward embryonic or FL-type hematopoiesis, and a deficiency of adult-type (i.e., bone marrow) hematopoiesis (20,21). In human development, embryonic, fetal, and adult-type hematopoiesis are characterized by expression of stage-specific β-like globins.…”

“…Human trisomy 21 pluripotent cells of various origins, human embryionic stem (hES), and induced pluripotent stem (iPS) cells, were differentiated in vitro as a model to recapitulate the effects of trisomy on hematopoiesis. To mitigate clonal variation, we isolated disomic and trisomic subclones from the same parental iPS line, thereby generating subclones isogenic except for chromosome 21. Under differentiation conditions favoring development of fetal liver-like, γ-globin expressing, definitive hematopoiesis, we found that trisomic cells of hES, iPS, or isogenic origins exhibited a two-to fivefold increase in a population of CD43 + (Leukosialin)/ CD235 + (Glycophorin A) hematopoietic cells, accompanied by increased multilineage colony-forming potential in colony-forming assays.…”

Multicolor Staining of Globin Subtypes Reveals Impaired Globin Switching During Erythropoiesis in Human Pluripotent Stem Cells

“…During erythropoiesis from hESCs and hiPSCs, the major globin forms are embryonic « and fetal g; little adult b-globin is expressed [7], indicating incomplete globin switching. Why the switching is not accomplished remains unclear, however [8][9][10][11][12][13][14][15][16][17][18].…”

Congenital amegakaryocytic thrombocytopenia iPS cells exhibit defective MPL-mediated signaling

“…Of importance in the present study is that MKs and erythrocytes generated in vitro from disease-specific iPSCs are an effective tool for studying the mechanism of not only thrombopoiesis (11), but also erythropoiesis (18)(19)(20).…”