Simultaeous detection of β‐galactosidase activity and surface antigen expression in viable haematopoietic cells

Berger, Christoph; Tan, Seong‐Seng; Sturm, Karin S.

doi:10.1002/cyto.990170305

Cited by 19 publications

(8 citation statements)

References 23 publications

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“…The following antibodies were used: CD45R/B220 (clone 6B2), CD3 (clone 145‐2C11), CD4 (clone RM4‐5), CD8 (clone 53‐6.7), TCRαβ (clone H57‐597) and TCRγδ (clone GL3), all from PharMingen (San Diego, CA). Intracellular β‐galactosidase activity was measured using the fluorogenic substrate fluorescein‐di‐β‐ D ‐galactopyranoside (FDG) in conjunction with flow cytometry (Berger et al ., 1994). In brief, 10 6 cells were incubated with a hypotonic solution containing 1 mM FDG for 50 s at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Activation of a novel proto-oncogene, Frat1, contributes to progression of mouse T-cell lymphomas

et al. 1997

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Acceleration of lymphomagenesis in oncogene‐bearing transgenic mice by slow‐transforming retroviruses has proven a valuable tool in identifying cooperating oncogenes. We have modified this protocol to search for genes that can collaborate effectively with the transgene in later stages of tumor development. Propagation of tumors induced by Moloney murine leukemia virus (M‐MuLV) in Eμ‐Pim1 or H2‐K‐myc transgenic mice by transplantation to syngeneic hosts permitted proviral tagging of ‘progression’ genes. Molecular cloning of common proviral insertion sites that were detected preferentially in transplanted tumors led to the identification of a novel gene, designated Frat1. The initial selection for integrations near Frat1 occurs in primary tumor cells that have already acquired proviruses in other common insertion sites, yielding primary lymphomas that contain only a minor fraction of tumor cells with an activated Frat1 allele. Transplantation of such primary lymphomas allows for a further expansion of tumor cell clones carrying a proviral insertion near Frat1, resulting in detectable Frat1 rearrangements in 17% of the transplanted Eμ‐Pim1 tumors and 30% of the transplanted H2‐K‐myc tumors, respectively. We have cloned and sequenced both the mouse Frat1 gene and its human counterpart. The proteins encoded by Frat1 and FRAT1 are highly homologous and their functions are thus far unknown. Tumor cell lines with high expression of Myc and Pim1 acquired an additional selective advantage in vivo upon infection with a Frat1‐IRES‐lacZ retrovirus, thus underscoring the role of Frat1 in tumor progression, and the ability of Frat1 to collaborate with Pim1 and Myc in lymphomagenesis.

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Section: Methodsmentioning

confidence: 99%

Activation of a novel proto-oncogene, Frat1, contributes to progression of mouse T-cell lymphomas

et al. 1997

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“…To determine ␤-galactosidase transgene expression, the cell populations were, prior to staining with monoclonal antibodies, incubated with fluorescein-di-␤-D-ga-51 Ts16 HEMATOPOIETIC DEVELOPMENT lactopyranoside as previously described (43). Expression of ␤-galactosidase in all the cells indicated male fetuses, while cell populations with about 50% ␤-galactosidase-positive cells indicated female fetuses.…”

Section: Flow Cytometrymentioning

confidence: 99%

Hematopoietic Deficiencies and Core Binding Factor Expression in Murine Ts16, an Animal Model for Down Syndrome

Gjertson

Sturm

Berger

1999

Clinical Immunology

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“…Cleavage of fluorescein di-o3-D-galactopyranosidase (FDG) was used to detect expression of LacZ [19,20]. To load cells with FDG, 50 Igl of a suspension containing 5 X 105 cells was prewarmed in a 37°C water bath for 10 min.…”

Section: Cell Suspensions and Flow Cytometrymentioning

confidence: 99%

Maternal Cells are Widely Distributed in Murine Fetuses in Utero1

Piotrowski

Croy

1996

Biology of Reproduction

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Passage of maternal cells into conceptuses in utero is recognized but poorly defined in species with hemochorial placentation. Despite the potential importance for such a phenomenon in vertical disease transmission, only limited data address the frequency of material to fetal cell trafficking or the developmental stage of its initiation. A murine model system, involving transfer of LacZ-, scid/scid, or wild type (+/+) blastocysts to pseudo-pregnant, LacZ+ transgenic ROSA26 females provided both flow cytometric and in situ information. In 100% of the late-gestation pregnancies studied, nucleated LacZ+ maternal cells crossed to conceptuses. In 90% of scid/scid fetuses, nucleated maternal cells were present in at least one lymphoid organ and often in more than one organ. Thymus was the most frequent site for maternal cell detection while the highest proportions of maternal cells were found in liver. Maternal cells were also visualized in fetal lung, heart, and bone marrow. Maternal cell trafficking into scid/scid fetuses commenced about midgestation, coincident with maturation of a placental circulation. In late-gestation +/+ fetuses, maternal cells were found extensively throughout bone marrow but not in other organs. The presence of maternal cells within primary lymphoid organs of fetuses may influence the repertoire of the developing fetal immune system and may be an underappreciated mechanism for vertical disease transmission.

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Simultaeous detection of β‐galactosidase activity and surface antigen expression in viable haematopoietic cells

Cited by 19 publications

References 23 publications

Activation of a novel proto-oncogene, Frat1, contributes to progression of mouse T-cell lymphomas

Activation of a novel proto-oncogene, Frat1, contributes to progression of mouse T-cell lymphomas

Hematopoietic Deficiencies and Core Binding Factor Expression in Murine Ts16, an Animal Model for Down Syndrome

Maternal Cells are Widely Distributed in Murine Fetuses in Utero1

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