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Updated: May 25, 2025
Pan-myeloid Differentiation of Human Cord Blood Derived CD34+ Hematopoietic Stem and Progenitor Cells
Published on: August 9, 2019
Distinct Phenotypic and Molecular Characteristics of CD34- and CD34+ Hematopoietic Stem/Progenitor Cell Subsets in
Ameera Gaafar1,2, Fatheia Nabeil Hamza1,2, Rama Yousif1
1Cell Therapy and Immunobiology Department, King Faisal Specialist Hospital and Research Center, P.O. Box 3354, Riyadh 11211, Saudi Arabia.
Background/Objectives: We aimed to identify the molecular signatures of primitive CD34+ and CD34- hematopoietic stem/progenitor cell (HSC/HPC) subsets in cord blood and bone marrow samples. Methods: CD34+ and CD34- HSC/HPC subsets from cord blood and bone marrow were characterized using flow cytometry, real-time PCR, and proteomic analysis to evaluate their phenotypic and molecular profiles. Results: Our findings revealed a significantly higher percentage of Lin-CD34-CD38Low/- (-/-) cells than of Lin-CD34+CD38Low/- (+/-) cells in cord blood. Aldehyde dehydrogenase levels were significantly lower in (-/-) than in (+/-) cells. Clonogenic ability was lower in (-/-) than in (+/-) cells. However, CD34- cells exhibited potent megakaryocyte/erythrocyte differentiation ability. Importantly, the HSC/HPC subsets expressed pluripotency or stemness genes (SOX2, Nanog, and OCT4); however, OCT4 expression significantly increased in (-/-) compared with (+/-) cells. We identified 304 proteins in the HSC/HPC subsets-85.6% had similar expression patterns in the two subsets; only 14.4% were differentially expressed between (-/-) and (+/-) cells. This implies their comparability at the protein level. Certain proteins were implicated in cellular-development-, gene-expression-, and embryonic-development-related signaling networks. Conclusions: Distinct biological and functional characteristics were observed between (-/-) and (+/-) HSC/HPC subsets. Some of the identified proteins may be novel HSC/HPC subsets markers for clinical applications after validation.
Background/Objectives: We aimed to identify the molecular signatures of primitive CD34+ and CD34- hematopoietic stem/progenitor cell (HSC/HPC) subsets in cord blood and bone marrow samples. Methods: CD34+ and CD34- HSC/HPC subsets from cord blood and bone marrow were characterized using flow cytometry, real-time PCR, and proteomic analysis to evaluate their phenotypic and molecular profiles. Results: Our findings revealed a significantly higher percentage of Lin-CD34-CD38Low/- (-/-) cells than of Lin-CD34+CD38Low/- (+/-) cells in cord blood. Aldehyde dehydrogenase levels were significantly lower in (-/-) than in (+/-) cells. Clonogenic ability was lower in (-/-) than in (+/-) cells. However, CD34- cells exhibited potent megakaryocyte/erythrocyte differentiation ability. Importantly, the HSC/HPC subsets expressed pluripotency or stemness genes (SOX2, Nanog, and OCT4); however, OCT4 expression significantly increased in (-/-) compared with (+/-) cells. We identified 304 proteins in the HSC/HPC subsets-85.6% had similar expression patterns in the two subsets; only 14.4% were differentially expressed between (-/-) and (+/-) cells. This implies their comparability at the protein level. Certain proteins were implicated in cellular-development-, gene-expression-, and embryonic-development-related signaling networks. Conclusions: Distinct biological and functional characteristics were observed between (-/-) and (+/-) HSC/HPC subsets. Some of the identified proteins may be novel HSC/HPC subsets markers for clinical applications after validation.
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