Distinct hematopoietic progenitor compartments are delineated by the expression of aldehyde dehydrogenase and CD34

Department of Surgery, Duke University, Durham, North Carolina, United States
Blood (Impact Factor: 10.45). 08/2005; 106(1):95-102. DOI: 10.1182/blood-2004-09-3652
Source: PubMed


A broad range of hematopoietic stem cells and progenitors reside within a fraction of umbilical cord blood (UCB) that exhibits low light scatter properties (SSC(lo)) and high expression of aldehyde dehydrogenase (ALDH(br)). Many SSC(lo) ALDH(br) cells coexpress CD34; however, other cells express either ALDH or CD34. To investigate the developmental potential of these cell subsets, purified ALDH(br) CD34+, ALDH(neg) CD34+, and ALDH(br) CD34(neg) UCB cells were characterized within a variety of in vivo and in vitro assays. Primitive progenitors capable of multilineage development were monitored in long- and short-term repopulation assays performed on nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice, and in primary and secondary long-term culture assays. These progenitors were highly enriched within the ALDH(br) CD34+ fraction. This cell fraction also enriched short-term myeloid progenitors that were detected in vitro. By comparison, ALDH(neg) CD34+ cells contained few primitive progenitors and had diminished short-term myeloid potential but exhibited enhanced short-term natural killer (NK) cell development in vitro. The ALDH(br) CD34(neg) cells were not efficiently supported by any of the assays used. These studies suggested that in particular the expression of ALDH delineated distinct CD34+ stem cell and progenitor compartments. The differential expression of ALDH may provide a means to explore normal and malignant processes associated with myeloid and lymphoid development.

Download full-text


Available from: Clay Smith,
  • Source
    • "CD34 þ cells suspended in 100 ml Aldefluor assay buffer and incubated for 30 min at 37°C (Storms et al., 2005). As a negative control, an aliquot of ALDF-stained cells was incubated with ALDH inhibitor diethylaminobenzaldehyde (DEAB). "
    [Show abstract] [Hide abstract]
    ABSTRACT: We analyzed the effect of exposure to hypoxic/hypercapnic (HH) gas mixture (5% O2 /9% CO2 ) on the maintenance of functional cord blood CD34(+) hematopoietic stem and progenitor cells in severe hypothermia (4°C) employing the physiological and proteomic approaches. Ten-day exposure to HH maintained the Day 0 (D-0) level of hematopoietic stem cells as detected in vivo on the basis of hematopoietic repopulation of immunodeficient mice-short-term scid repopulating cells (SRC). Conversely, in the atmospheric air (20% O2 /0.05% CO2 ), usual condition used for cell storage at 4°C, stem cell activity was significantly decreased. Also, HH doubled the survival of CD34(+) cells and committed progenitors (CFCs) with respect to the atmospheric air (60% vs. 30%, respectively). Improved cell maintenance in HH was associated with higher proportion of aldehyde dehydrogenase (ALDH) positive cells. Cell-protective effects are associated with an improved maintenance of the plasma and mitochondrial membrane potential and with a conversion to the glycolytic energetic state. We also showed that HH decreased apoptosis, despite a sustained ROS production and a drop of ATP amount per viable cell. The proteomic study revealed that the global protein content was better preserved in HH. This analysis identified: (i) proteins sensitive or insensitive to hypothermia irrespective of the gas phase, and (ii) proteins related to the HH cell-protective effect. Among them are some protein families known to be implicated in the prolonged survival of hibernating animals in hypothermia. These findings suggest a way to optimize short-term cell conservation without freezing. © 2014 Wiley Periodicals, Inc.
    Journal of Cellular Physiology 12/2014; 229(12). DOI:10.1002/jcp.24678 · 3.84 Impact Factor
  • Source
    • "The leukocyte fraction (1 × 106 WBC/mL) was prepared by adding assay buffer (Aldagen, Durham, CA, USA) to the cell pellet. Next, 0.5 mL of the leukocyte fraction was incubated with 5 μL of the fluorescent reagent BAAA, which is specific for the aldehyde dehydrogenase expressed in highly immature precursors of leukocytes, for 30–60 minutes at 37°C [10, 18]. After incubation, the mixture was centrifuged at 250 ×g for 5 minutes, the supernatant was removed, the cell pellet was resuspended in 0.5 mL of assay buffer (Aldagen, Durham, CA, USA), and the mixture was analysed immediately. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Hematopoietic stem cells (HSCs), still represent a certain mystery in biology, have a unique property of dividing into equal cells and repopulating the hematopoietic tissue. This potential enables their use in transplantation treatments. The quality of the HSC grafts for transplantation is evaluated by flow cytometric determination of the CD34(+) cells, which enables optimal timing of the first apheresis and the acquisition of maximal yield of the peripheral blood stem cells (PBSCs). To identify a more efficient method for evaluating CD34(+) cells, we compared the following alternative methods with the reference method: hematopoietic progenitor cells (HPC) enumeration (using the Sysmex XE-2100 analyser), detection of CD133(+) cells, and quantification of aldehyde dehydrogenase activity in the PBSCs. 266 aphereses (84 patients) were evaluated. In the preapheretic blood, the new methods produced data that were in agreement with the reference method. The ROC curves have shown that for the first-day apheresis target, the optimal predictive cut-off value was 0.032 cells/mL for the HPC method (sensitivity 73.4%, specificity 69.3%). HPC method exhibited a definite practical superiority as compared to other methods tested. HPC enumeration could serve as a supplementary method for the optimal timing of the first apheresis; it is simple, rapid, and cheap.
    BioMed Research International 09/2014; 2014:575368. DOI:10.1155/2014/575368 · 2.71 Impact Factor
  • Source
    • "ALDH is highly expressed in primitive hematopoietic cells, and surface markers reflecting hematopoietic stem cell activity, such as CD34 and CD133, have been reported to be expressed on some fraction of ALDHbr cells [22]. According to previous studies, CD34+ALDHbr cells and ALDHbrCD34+ cells affect long-term xenograft and multi-lineage hematopoiesis [22,23, 24]. Apoptotic CD34+ cells have been reported to be able to influence transplantation outcome [25]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Although cord blood (CB) is a well-known source of hematopoietic stem cells, uncertainties exist regarding the quality of cryopreserved CB. We investigated the changes in quality of CB units according to the duration of cryopreservation. We analyzed CB units that were rejected from the Seoul Metropolitan Government Public Cord Blood Bank inventory after conventional processing, because of unsuitability for allogeneic transplantation. Two hundred CB units that were cryopreserved from 1 year to 5 years were selected. After thawing the cryopreserved CB units, the total nucleated cell (TNC) count, CD34+ cell count, number of colony-forming units (CFU), aldehyde dehydrogenase (ALDH) level, cell viability, and apoptosis were analyzed. We conducted a comparative analysis to identify the presence of statistically significant differences in the recovery rates of the TNC and CD34+ cell counts and to compare the results of ALDH level, the cell viability test, the apoptosis test, and CFU analysis among groups according to the duration of cryopreservation. The recovery rates of the TNC count, the CD34+ cell count, and cell viability did not differ significantly according to the duration of cryopreservation. ALDH analysis, the cell viability test, and the apoptosis test did not reveal any increasing or decreasing trend according to the duration of cryopreservation. Further, the numbers of CFU-granulocyte/macrophage and CFU-granulocyte/erythrocyte/macrophage/megakaryocyte did not differ significantly according to the duration of cryopreservation. These results suggest that the quality of CB is not affected by cryopreservation for up to a period of 5 years.
    Blood Research 03/2014; 49(1):54-60. DOI:10.5045/br.2014.49.1.54
Show more