Polychromatic flow cytometry analysis of CD34+ hematopoietic stem cells in cryopreserved early preterm human cord blood samples

Abstract

During the last decades, extended characterizations were performed of human fullterm cord blood (hTCB) cells, but little information is available on human early preterm cord blood (hEPCB) hematopoietic stem cells (HSCs). In our study, we analyzed by flow cytometry 19 hEPCB and 17 hTCB samples. First, we observed that the percentage of CD34 Pos CD45 Dim cells was higher in hEPCB compared with hTCB and that it decreased during 16th-20th week of pregnancy. Within the CD34 Pos CD45 Dim population, we examined the expression of CD29, CD31, CD38, CD90, CD117, CD133, CD135, CD200, CD243, and CD338. We found that CD135 intensity and CD243 Pos cells percentage were lower in hEPCB compared with hTCB. As to CD38, we observed that hEPCB samples were richer in undifferentiated CD34Pos CD45 Dim CD38 Neg HSCs compared with hTCB counterparts. We also compared the expression of the abovementioned molecules in undifferentiated and committed HSCs residing in hEPCB and hTCB. CD133Pos cells compared with hTCB samples. Finally, analyzing monocytes and lymphocytes within the two samples, we observed that T-cell percentages were higher in hTCB, whereas B-cell percentages were higher in hEPCB. We, therefore, studied the B-cell lineage maturation and found a higher percent of pro-B and pre-B cells in hEPCB compared with hTCB samples. Taken together, these results evidence the hematopoietic peculiarity of hEPCB, potentially useful for highlighting early steps of human hematolymphopoiesis as well as for developing novel strategies of stem cell-based therapy. ' 2010 International Society for Advancement of Cytometry Key termsfetal cord blood; CD34; hematopoietic stem cells; flow cytometry; B-lymphocytes; early preterm cord blood DURING the past decade, human full-term cord blood (HTCB) has been considered to be an attractive hematopoietic stem cells (HSC) source not only for transplantation purposes, but also for basic research investigators studying useful marker to identify immature stages of HSC differentiation (1-3). Compared with mobilized peripheral blood stem cells and marrow HSCs, CB-HSCs are essentially characterized by a series of advantages such as less-stringent HLA-matching requirements between donors and recipients, absence of ethical concerns as well as risks for the mother and the infant, low Epstein Barr virus and cytomegalovirus contaminations, immediate availability of frozen units, and maintenance of ethnic balance by targeted collection programs (1).

Micro-Computed Tomography Detection of Gold Nanoparticle-Labelled Mesenchymal Stem Cells in the Rat Subretinal Layer

“…The MSCs were successfully culture-expanded from human Wharton’s jelly (hWJ-MSCs) (Figure 1) and were shown to be multipotent, since the cells were able to differentiate into adipocytes and osteocytes, which are both part of the mesoderm lineage (Figure 2). These cells highly expressed CD73, CD90, and CD105, but lacked expression of CD14, CD34, and CD45 (Figure 3), indicating the absence of contaminating hematopoietic cells in the culture [22]. In accordance with previous reports, these cells also demonstrated a lack of the co-stimulatory molecules CD80 and CD86 [23], conferring the ability to escape from immune surveillance.…”

Phenotypic and functional characterization of cytokine-induced killer cells derived from preterm and term infant cord blood

“…The results showed that PCB-CIK cells had a higher proliferation rate than the TCB-CIK cells on days 7, 8, 9 and 10. The possible cause is that preterm infant cord blood has more immature and undifferentiated cells (22). However, the PCB-CIK cells were difficult to cultivate.…”

Human adult stem cells from diverse origins: An overview from multiparametric immunophenotyping to clinical applications

“…Recently published manuscripts, summarize flow cytometry characterization and specification of these stem cell based on phenotypic and differentiation stage‐specific markers, for isolation and studying their biology and clinical applications . Multiparametric flow cytometry and image stream analysis, the latter one combining morphological and imaging analysis with population‐relevant antigene expression are being used for identifying tissue‐specific stem cells, as well as for tumor cells and subpopulations within them in humans and animals .…”