التفاصيل البيبلوغرافية
| العنوان: |
Definitive hematopoietic stem/progenitor cells from human embryonic stem cells through serum/feeder-free organoid-induced differentiation. |
| المؤلفون: |
Demirci, Selami, Haro-Mora, Juan J., Leonard, Alexis, Drysdale, Claire, Malide, Daniela, Keyvanfar, Keyvan, Essawi, Khaled, Vizcardo, Raul, Tamaoki, Naritaka, Restifo, Nicholas P., Tisdale, John F., Uchida, Naoya |
| المصدر: |
Stem Cell Research & Therapy; 11/24/2020, Vol. 11 Issue 1, pN.PAG-N.PAG, 1p |
| مصطلحات موضوعية: |
EMBRYONIC stem cells, HUMAN embryonic stem cells, PROGENITOR cells, HEMATOPOIETIC stem cells, T cell receptors |
| مستخلص: |
Background: Ex vivo production of hematopoietic stem/precursor cells (HSPCs) represents a promising versatile approach for blood disorders. Methods: To derive definitive HSPCs from human embryonic stem cells (ESCs), we differentiated mesodermally specified embryoid bodies (EBs) on gelatin-coated plates in serum/feeder-free conditions. Results: Seven-day EB maturation followed by an 8-day differentiation period on OP9 cells provided the highest number of definitive (CD34+ CD235a−, 69%, p < 0.01) and lowest number of primitive (CD34− CD235a+, 1.55%, p < 0.01) precursor cells along with the highest colony-forming units (149.8 ± 11.6, p < 0.01) in feeder-free conditions. Maximal HSPC fraction (CD34+ CD38− CD45RA− CD49f+ CD90+) was 7.6–8.9% after 10 days of hematopoietic differentiation with 14.5% adult β-globin expression following RBC differentiation. Myeloid and erythroid colonies were restricted strictly to the CD34+ CD43+ fraction (370.5 ± 65.7, p < 0.001), while the CD34− CD43+ fraction produced only a small number of colonies (21.6 ± 11.9). In addition, we differentiated the CD34+ CD43+ cells towards T-lymphocytes using the OP9/DLL1 co-culture system demonstrating double-positive T cells (CD4+ CD8+) with CD3+ expression displaying a broad T cell receptor (TCR) repertoire. Confocal imaging of organoid-like structures revealed a close association of CD31+ cells with CD34+ and CD43+ cells, suggesting a potential emergence of HSPCs through endothelial to hematopoietic transition. Furthermore, fluorescently labeled organoids exhibited the emergence of spherical non-attached cells from rare progenitors at the border of the organoid center. Conclusions: In summary, definitive HSPCs can be derived from ESCs through a dynamic cellular process from an organoid-like structure, where erythroid progeny are capable of producing adult hemoglobin and lymphoid progeny shows a diverse TCR repertoire. [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
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