Circulating hematopoietic progenitor cells in a fetus with alpha thalassemia: comparison with the cells circulating in normal and non-thalassemic anemia fetuses and implications for in utero transplantations

Abstract

Summary:Our aim was to evaluate the number of progenitor cells circulating in an ␣-thalassemic fetus during its infusion in utero with paternal CD34 + and adult red cells and to compare those values with those circulating in normal and non-thalassemic anemic fetuses of matched gestational age. The treatment of the ␣-thalassemic fetus has been described elsewhere. Fetal blood was obtained from normal and anemic fetuses by fetal blood sampling for diagnostic or therapeutic purposes according to a protocol approved by the human subject committee. The number of progenitor cells in fetal blood was estimated on the basis of the number of colonies they gave rise to in semisolid cultures. The ␣-thalassemic fetus, as did the other fetuses analyzed, contained high numbers (10 6 -10 7 depending on the age) of progenitor cells, values which were higher than the number (10 4 -10 5 ) of paternal progenitor cells being transplanted. Progenitor cells with adult characteristics (adult kinetics of differentiation) were detected rapidly (10 min) after the CD34 + cell infusion, but were not detectable 2-3 weeks after the transplant. These results indicate that adult progenitor cells do not have a numerical advantage when transplanted into ␣-thalassemic fetuses.

Identification of Biologically Relevant Enhancers in Human Erythroid Cells

“…Cell Culture and Selection-To obtain primary human erythroid cells, CD34 ϩ cells were cultured and selected as described (41,42). These cells represent the R3/R4 cell population of nucleated erythroid cells defined by Zhang et al (43).…”

Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis

“…K562 cells (chronic myelogenous leukemia in blast crisis with erythroid characteristics, ATCC catalog CCL-243) were maintained in RPMI 1640 medium with 10% fetal calf serum. Human CD34-selected stem and progenitor cells were obtained from the Yale Center for Excellence in Molecular Hematology Cell Core and cultured in StemSpan SF expansion medium (StemSpan, 09650) with estradiol (100 ng/ml), dexamethasone (10 ng/ml), human transferrin (200 ng/ml), insulin (10 ng/ml), Flt3 ligand (100 n/ml), stem cell factor (100 ng/ml), IL-3 (50 ng/ml), IL-6 (20 ng/ml), insulin like growth factor-1 (50 ng/ml), and erythropoietin (3 U/ml) for 9-14 days (85,86). FACS analysis was used to analyze cellular expression of CD71 (transferrin receptor) and CD235a (glycophorin A).…”

Erythrocyte peripheral type benzodiazepine receptor/voltage-dependent anion channels are upregulated by Plasmodium falciparum

“…Human mobilized peripheral blood CD34-selected stem and progenitor cells were obtained from the Yale Center of Excellence in Molecular Hematology Cell Core and cultured in StemSpan SF expansion medium (StemSpan 09650) with estradiol (100 ng/mL), dexamethasone (10 ng/mL), human transferrin (200 ng/mL), insulin (10 ng/mL), Flt3 ligand (100 ng/mL), stem cell factor (100 ng/mL), IL-3 (50 ng/mL), IL-6 (20 ng/mL), insulin-like growth factor 1 (50 ng/mL), and erythropoietin (3 U/mL) for 9 to 14 days. 13,14 FACS analysis was used to analyze the cellular expression of CD71 (transferrin receptor) and CD235a (glycophorin A). Magnetic bead selection for CD71 (MACS 130-046-201; Miltenyi Biotech) and CD235a (MACS 130-050-501; Miltenyi Biotech) was used to purify an R3/R4 population of erythroid cells.…”