Distinct Phenotypic and Molecular Characteristics of CD34- and CD34+ Hematopoietic Stem/Progenitor Cell Subsets in Cord Blood and Bone Marrow Samples: Implications for Clinical Applications.

PMID 40002599 | PMCID PMC11853955 | DOI 10.3390/diagnostics15040447 · Diagnostics (Basel, Switzerland) · 2025

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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.