RETRACTED ARTICLE: Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells

Huang, Shiang; Terstappen, Leon W.M.M.

doi:10.1038/360745a0

Cited by 149 publications

(68 citation statements)

References 16 publications

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“…Single cells were sorted into 96-well dishes (Costar; Cambridge, MA) using FACStar plus with an automated cell deposition unit. The accuracy of single-cell sorting and deposition is estimated to be >99% [44]. A typical sort window for CD34 + cells with different levels of CD123 expression is shown in Figure 1.…”

Section: Flow Cytometry and Cell Sortingmentioning

confidence: 99%

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues

Huang¹,

Chen²,

Yu³

et al. 1999

STEM CELLS

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CD123 (␣-subunit of IL-3 receptor) expression on primitive and committed human hematopoietic cells was studied by multicolor sorting and single-cell culture. The sources of cells included fetal liver (FLV), fetal bone marrow, umbilical cord blood, adult bone marrow and mobilized peripheral blood. Three subsets of CD34 + cells were defined by the levels of surface CD123: CD123 negative , CD123 low , and CD123 bright . Coexpression of lineage markers showed that a majority of CD34 + CD123 bright cells were myeloid and B-lymphoid progenitors, while erythroid progenitors were mainly in the CD34 + CD123 negative subset. The CD34 + CD123 low subset contained a heterogeneous distribution of early and committed progenitor cells. Single CD34 + cells from the CD123 subsets were cultured in a cytokine cocktail of stem cell factor, interleukin 3 (IL-3), IL-6, GM-CSF, erythropoietin, insulin-like growth factor-1, and basic fibroblast growth factor. After 14 days of incubation, a higher cloning efficiency (CE) was observed in the CD34 + CD123 negative and CD34 + CD123 low fractions (37 ± 23% and 44 ± 23%, respectively) than in the CD34 + CD123 bright fraction (15 ± 21%). Using previously published criteria that colonies containing dispersed, translucent cells (dispersed growth pattern, DGP) were derived from primitive cells and that colonies composed solely of clusters were from committed cells, early precursors were distributed evenly in the CD34 + CD123 negative and CD34 + CD123 low subsets. When CD38 and CD90 (Thy-1) were used for further characterization of CD34 + cells from FLV, CE increased from 37 ± 23% in CD123 negative to 70 ± 19% in CD123 negative CD38 -and from 44 ± 23% in CD123 low to 66 ± 19% in CD123 low CD38 -. No significant increase in CE or DGP progenitors was observed when CD34 + cells were sorted by CD90 and CD123. We concluded that: A) high levels of CD123 were expressed on B-lymphoid and myeloid progenitors; B) early erythroid progenitors had little or no surface CD123, and C) primitive hematopoietic cells are characterized by CD123 negative/low expression.

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Section: Flow Cytometry and Cell Sortingmentioning

confidence: 99%

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues

Huang¹,

Chen²,

Yu³

et al. 1999

STEM CELLS

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“…Cells were analyzed by flow cytometry to assess the proportion of CD34 + cells. 20 Vector production and transduction Retroviral vector LNL and the anti-HIV ribozyme expression vector were made by transfection of corresponding plasmids into a PA317 package cell line. Colonies secreting high titer of retroviral particles were selected and cultured.…”

Section: Enrichment Of Cd34 + Cellsmentioning

confidence: 99%

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

Gervaix

Kang

et al. 1998

Gene Ther

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Hematopoietic CD34+ cells from placental and umbilical cord blood (PUCB) can be valuable vehicles for gene therapy of immunodeficiencies and genetic disorders. We have conducted preclinical studies towards the treatment of HIV-1-infected infants with genetically 'immunized' CD34+ cells derived from PUCB using anti-HIV-1 hairpin ribozyme genes. PUCB was collected from 10 newborns of HIV-1-positive mothers. CD34+ cells were enriched with a modified procedure using Dynal immunomagnetic beads and chymopapain, stimulated with stem cell factor (SCF), IL-3 and IL-6, and transduced using cell-free recombinant retroviral vector (MJT) expressing a ribozyme against the U5 region of HIV-1. No significant differences were observed in the growth pattern of CD34+ cells from normal infants, HIV-1 exposed infants or infants confirmed to be infected by HIV-1. The transduction efficiency of the CD34+ cells from all the infants was also comparable. MJT-transduced CD34+ cells from an HIV-1-infected infant were maintained in a liquid culture system for 4 weeks, and the progeny macrophage cells were challenged with the monocyte-tropic laboratory strain, HIV-Bal, or the HIV-1 isolate from the infant's mother. Significant inhibition of virus replication was observed in ribozyme-transduced cells. Thus, we have demonstrated efficient and stable gene transfer into progenitor cells from the cord blood of HIV-1-exposed or -infected infants and shown that protection from HIV-1 infection was conferred to the progeny cells produced by CD34+ cells transduced with the anti-HIV ribozyme gene construct

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“…The identification of a cell which can give rise in single cell cultures to both mesenchymal elements ("stroma") and primitive leukocytes has recently been reported [31]. These cells, which were isolated from fetal bone marrow, express the CD34 antigen but do not appear to express lineage-associated antigens or the activation markers CD38 and HLA-DR.…”

Section: Hematopoietic Stem Cellsmentioning

confidence: 99%

CD34 antigen: Molecular features and potential clinical applications

1993

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Despite the wide variety of functions exhibited by mature peripheral blood cells, all are derived from a small pool (1–3%) of primitive precursor cells in the bone marrow (BM) that bear a unique surface glycoprotein, CD34. Isolated CD34+ cells are capable of reconstituting all hematopoietic lineages, both in experimental animals and in humans following intensive therapy. CD34+ cells capable of reconstituting hematopoiesis are also found at low frequency in peripheral blood (PB), a frequency which can be dramatically increased by combinations of chemotherapy and recombinant cytokines. In some cases, PB “stem cells” (PBSC) can be used to augment or even replace conventional BM autografts. The availability of CD34 antibodies has greatly aided the development of techniques for the enrichment of primitive progenitor cells, thus allowing studies of the hematopoietic potential of stem cells in vitro. Additionally, the use of CD34 antibodies for the “positive selection” of hematopoietic stem/progenitor cells from tumor‐contaminated marrow may possibly represent an alternative “purging” strategy prior to transplantation. The availability of pure populations of the most primitive hematopoietic progenitor cells will also facilitate study of genetic manipulation as a practical therapeutic modality.

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RETRACTED ARTICLE: Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells

Cited by 149 publications

References 16 publications

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

CD34 antigen: Molecular features and potential clinical applications

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RETRACTED ARTICLE: Formation of haematopoietic microenvironment and haematopoietic stem cells from single human bone marrow stem cells

Cited by 149 publications

References 16 publications

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 + Hematopoietic Cells from Different Tissues

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 + Hematopoietic Cells from Different Tissues

Gene therapy targeting cord blood-derived CD34+ cells from HIV-exposed infants: preclinical studies

CD34 antigen: Molecular features and potential clinical applications

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Product

Resources

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Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues

Correlation Between IL‐3 Receptor Expression and Growth Potential of Human CD34 ⁺ Hematopoietic Cells from Different Tissues