Morphological characterization of stromal cell types in hematopoietically                active long-term murine bone marrow cultures.

Hauser, Simon P.; Waldron, James A.; Upuda, K B; Lipschitz, David A.

doi:10.1177/43.4.7897180

Cited by 13 publications

(13 citation statements)

References 8 publications

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“…The human BM stromal cell population appeared to consist predominantly of fibroblastic cells whereas no vWF-positive endothelial-like cells could be detected in accordance with previous observations (Bentley and Foidart, 1980;Bentley and Tralka 1982), although other reports indicated that endotheliallike cells could be isolated from long-term BM cultures (Zuckerman and Wicha, 1983;Fei et al, 1990;Hauser et al, 1995). These differences may be due to culture conditions and/or kinetic differences of cell growth and differentiation.…”

Section: Discussionsupporting

confidence: 88%

Bone Marrow Stromal Cells as Targets for Gene Therapy of Hemophilia A

Chuah¹,

Brems²,

Vanslembrouck³

et al. 1998

Human Gene Therapy

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Attempts to develop an ex vivo gene therapy strategy for hemophilia A, using either primary T cells or bone marrow (BM) stem/progenitor cells have been unsuccessful, due to the inability of these cell types to express coagulation factor VIII (FVIII). As an alternative, we evaluated the potential of BM-derived stromal cells which can be readily obtained and expanded in vitro. Human and murine BM stromal cells were transduced with an intron-based Moloney murine leukemia virus (MoMLV) retroviral vector expressing a B-domain-deleted human factor VIII cDNA (designated as MFG-FVIIIdeltaB). Transduction efficiencies were increased 10- to 15-fold by phosphate depletion and centrifugation, which obviated the need for selective enrichment of the transduced BM stromal cells. This resulted in high FVIII expression levels in transduced human (180 +/- 4 ng FVIII/10[6] cells per 24 hr) and mouse (900 +/- 130 ng FVIII/10[6] cells per 24 hr) BM stromal cells. Pseudotyping of the MFG-FVIIIdeltaB retroviral vectors with the gibbon ape leukemia virus envelope (GALV-env) resulted in significantly higher transduction efficiencies (100 +/- 20%) and FVIII expression levels (390 +/- 10 ng FVIII/10[6] cells per 24 hr) in transduced human BM stromal cells than with standard amphotropic vectors. This difference in transduction efficiency correlated with the higher titer of the GALV-env pseudotyped viral vectors and with the higher GALV receptor (GLVR-1) versus amphotropic receptor (GLVR-2) mRNA expression levels in human BM stromal cells. These findings demonstrate the potential of BM stromal cells for gene therapy in general and hemophilia A in particular.

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Section: Discussionsupporting

confidence: 88%

Bone Marrow Stromal Cells as Targets for Gene Therapy of Hemophilia A

Chuah¹,

Brems²,

Vanslembrouck³

et al. 1998

Human Gene Therapy

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“…42 In addition, antagonism by serotonin of interferon g via 5HT2 receptors in the marrow macrophage has been demonstrated. 43 The present study found 5ht2a mRNA in stromal cells of the marrow which contain macrophages, 44 although cell-type-specific localization was not determined.…”

Section: Discussioncontrasting

confidence: 50%

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

et al. 2003

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The antipsychotic drug clozapine, acts via interaction with selective neurotransmitter receptor systems. Its use however, is associated with lifethreatening agranulocytosis. The mechanism by which this occurs and its possible relationship with the drug's atypicality remain unclear. As a first step in identifying mechanistic pathways involved, profiling of neurotransmitter receptors on human neutrophils, mononuclear and bone marrow stromal cells as putative targets for clozapine-mediated toxicity was undertaken. Expression of mRNA encoding dopaminergic d2, d3, d4; serotonergic 5ht2a, 5ht2c, 5ht3, 5ht6, 5ht7; adrenergic a1a, a2; histaminergic h1 and muscarinic m1, m2, m3, m4, m5 receptors was analyzed by reverse transcription-polymerase chain reaction methods. While 5ht2c, 5ht6, m1 and m2 mRNA were undetected, the presence of the other receptors indicates sites at which clozapine could bind and induce toxicity of neutrophils and stromal components which regulate granulopoiesis. The functional significance of differential receptor expression while unknown, may argue for neural regulation of hematopoiesis.

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“…The morphology of these cells, shown in Figure 1, endothelial cells have been reported earlier. 12 Investigation for the presence of primitive hematopoietic cells showed no c-kit ligand receptor positive cells on flow cytometric analysis confirming lack of contamination by hematopoietic progenitor cells (data not shown).…”

Section: Resultsmentioning

confidence: 72%

“…12,28 Briefly, 10 7 cells, resuspended in 10 ml of the medium described above, were plated in 100-mm dishes and incubated at 37°C in 5% CO 2 . At weekly intervals the cultures were fed by replacing 50% of the supernatant with fresh culture medium.…”

Section: Stromal Cell Culturementioning

confidence: 99%