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A promising and increasingly exploited property of hematopoietic stem cells is their ability to efflux the fluorescent dye Hoechst 33342. The Hoechst-negative cells are isolated by fluorescence-activated cell sorting as a so-called side "population" (SP) of bone marrow. This SP from bone marrow, as well as other tissues, is reported to contain immature stem(More)
The PM27 gene encodes a sea urchin skeletal protein. Both the transcript and encoded protein appear at the mesenchyme blastula stage and are restricted to the primary mesenchyme cell (PMC) lineage throughout development. Transgenic expression of PM27 promoter constructs demonstrates that this cell specificity is regulated at the level of transcription. The(More)
In order to examine the ontogeny of tissue-specific expression of the msp130 gene during early embryogenesis of the sea urchin, we have developed a whole-mount, non-radioactive in situ hybridization protocol suitable for these embryos. This protocol is adapted from the existing technology of immunohistochemical localization of digoxygenin-labelled(More)
In this report we identify a 130-kDa protein encoded by a sea urchin primary mesenchyme-specific cDNA clone, 18C6. The cDNA clone has been partially sequenced, and an open reading frame has been identified. A portion of this open reading frame has been expressed as a beta-galactosidase fusion protein in Escherichia coli, and antibodies to the fusion protein(More)
Methods are described for isolation and culture of primary mesenchyme cells from echinoid embryos. Ninety-five percentpure primary mesenchyme cells were isolated from early gastrulae ofStrongylocentrotus purpuratus, exploiting the biological segregation of these cells within the blastocoel. When cultured, more than 90% of the isolated cells reached the(More)
Retroviral integration provides a unique and heritable genomic tag for a target cell and its progeny, enabling studies of clonal composition and repopulation kinetics after gene transfer into hematopoietic stem cells. The clonal tracking method, linear amplification-mediated polymerase chain reaction (LAM-PCR) is widely employed to follow the hematopoietic(More)
Canine embryonic stem (cES) cell lines were generated to establish a large-animal preclinical model for testing the safety and efficacy of embryonic stem (ES) cell-derived tissue replacement therapy. Putative cES cell lines were initiated from canine blastocysts harvested from natural matings. Times of harvest were estimated as 12-16 days after the presumed(More)
Major histocompatibility complex (MHC) genes in mammals include highly polymorphic class I and class II genes that are critical for donor-recipient matching for transplantation. Dogs have served as an effective, directly translatable model for stem/progenitor cell transplantation. Previous analyses of MHC class I genes in dogs point to a single highly(More)
The crystals found in sporulation extracts of Bacillus thuringiensis (Berliner) contain proteins that are highly toxic to insects. Different crystal proteins exhibit distinct specificities for restricted groups of insects. An uncharacterized strain of B. thuringiensis (BtS2), derived from China, was found to carry several crystal protein genes and to be(More)
Gene therapy for hemoglobinopathies requires efficient gene transfer into hematopoietic stem cells and high-level erythroid-specific gene expression. Toward this goal, we constructed a helper-dependent adenovirus vector carrying the beta-globin locus control region (LCR) to drive green fluorescent protein (GFP) expression, whereby the LCR-GFP cassette is(More)