scholarly journals FADD and caspase-8 are required for cytokine-induced proliferation of hemopoietic progenitor cells

Blood ◽  
2005 ◽  
Vol 106 (5) ◽  
pp. 1581-1589 ◽  
Author(s):  
Marc Pellegrini ◽  
Sue Bath ◽  
Vanessa S. Marsden ◽  
David C. S. Huang ◽  
Donald Metcalf ◽  
...  
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Caspase 8 ◽  
Liver Stem Cells ◽  
Hemopoietic Progenitor ◽  
Hemopoietic Progenitor Cells ◽  
Negative Mutant

Abstract The role of caspase-8 and its adaptor Fas-associated death domain (FADD) in lymphocyte apoptosis is well defined, but their functions in other hemopoietic lineages are not clear. We were unable to generate transgenic mice expressing dominant inhibitors of FADD or caspase-8 in hemopoietic cells, possibly because their expression may have precluded production of vital hemopoietic cells. When using a retroviral gene delivery system, fetal liver stem cells expressing a dominant-negative mutant of FADD (FADD-DN) were unable to generate myeloid or lymphoid cells upon transplantation into lethally irradiated mice. However, fetal liver stem cells expressing very low levels of the caspase-8 inhibitor cytokine response modifier A (CrmA) could reconstitute the hemopoietic system. This level of CrmA expression provided some protection against Fas ligand (FasL)–induced apoptosis and promoted accumulation of myeloid cells in the bone marrow, but it did not inhibit mitogen-induced proliferation of B or T lymphocytes. Using an in vitro colony formation assay, we found that fetal liver stem cells expressing FADD-DN, CrmA, or a dominant-negative mutant of caspase-8 could not proliferate in response to cytokine stimulation. These data demonstrate that the enzymatic activity of caspase-8 and its adaptor FADD are required for cytokine-induced proliferation of hemopoietic progenitor cells.

Hepatic stem cells and their enzymatic markers- A review

2018 ◽  
Author(s):  
Amit Kumar ◽  
Sourabh Sulabh
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Lactate Dehydrogenase ◽  
Progenitor Cells ◽  
Ethical Issues ◽  
Fetal Liver ◽  
Lactate Dehydrogenase Activity ◽  
Liver Stem Cells ◽  
Enzymatic Markers

Stem cells are those cells which show capacity for self-renewal and ability to give rise to multiple differentiated cellular populations. Enzymatic activity, as a marker for cell proliferation and cell viability, is used by metabolic activity assays. Liver stem cells/progenitor cells can be a useful source of liver treatment. They can repopulate and restore injured liver. Fetal liver stem/ progenitor cells have been found to be more capable in this, but are subjected to ethical issues. Adult liver stem cells and stem cells from animals can be used. Alkaline phosphatase and lactate dehydrogenase are enzymatic markers of in vitro hepatocyte culture. During in vitro cell culture, in the culture medium, secreted alkaline phosphatase activity increases during exponential growth of cells, whereas low extracellular lactate dehydrogenase activity indicates increased number of viable cells. Alkaline phosphatase and lactate dehydrogenase activities can be used to assess hepatocytes proliferation in vitro.

scholarly journals Discovering and Characterizing of Survivin Dominant Negative Mutants With Stronger Pro-apoptotic Activity on Cancer Cells and CSCs

2021 ◽  
Vol 11 ◽  
Author(s):  
Wei Guo ◽  
Xingyuan Ma ◽  
Yunhui Fu ◽  
Chang Liu ◽  
Qiuli Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cancer Cells ◽  
A549 Cells ◽  
Dominant Negative ◽  
Dominant Negative Mutant ◽  
Anti Cancer ◽  
Apoptotic Activity ◽  
Reversal Index ◽  
Negative Mutant

Survivin as a member of the inhibitor of apoptosis proteins (IAPs) family is undetectable in normal cells, but highly expressed in cancer cells and cancer stem cells (CSCs) which makes it an attractive target in cancer therapy. Survivin dominant negative mutants have been reported as competitive inhibitors of endogenous survivin protein in cancer cells. However, there is a lack of systematic comparative studies on which mutants have stronger effect on promoting apoptosis in cancer cells, which will hinder the development of novel anti-cancer drugs. Here, based on the previous study of survivin and its analysis of the relationship between structure and function, we designed and constructed a series of different amino acid mutants from survivin (TmSm34, TmSm48, TmSm84, TmSm34/48, TmSm34/84, and TmSm34/48/84) fused cell-permeable peptide TATm at the N-terminus, and a dominant negative mutant TmSm34/84 with stronger pro-apoptotic activity was selected and evaluated systematically in vitro. The double-site mutant of survivin (TmSm34/84) showed more robust pro-apoptotic activity against A549 cells than others, and could reverse the resistance of A549 CSCs to adriamycin (ADM) (reversal index up to 7.01) by decreasing the expression levels of survivin, P-gp, and Bcl-2 while increasing cleaved caspase-3 in CSCs. This study indicated the selected survivin dominant negative mutant TmSm34/84 is promising to be an excellent candidate for recombinant anti-cancer protein by promoting apoptosis of cancer cells and their stem cells and sensitizing chemotherapeutic drugs.

Expression and function of c-Kit in fetal hemopoietic progenitor cells: transition from the early c-Kit-independent to the late c-Kit-dependent wave of hemopoiesis in the murine embryo

Development ◽  
1993 ◽  
Vol 117 (3) ◽  
pp. 1089-1098 ◽  
Author(s):  
M. Ogawa ◽  
S. Nishikawa ◽  
K. Yoshinaga ◽  
S. Hayashi ◽  
T. Kunisada ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Progenitor Cells ◽  
Fetal Liver ◽  
Flow Cytometric Analysis ◽  
Yolk Sac ◽  
Erythroid Progenitor ◽  
Erythroid Progenitor Cells ◽  
And Function ◽  
Hemopoietic Progenitor ◽  
Hemopoietic Progenitor Cells

The protooncogene c-kit encodes a receptor type tyrosine kinase and is allelic with the W locus of mice. SLF, the c-Kit ligand which is encoded by the Sl locus, has growth promoting activity for hemopoietic stem cells. Previous studies demonstrated that c-Kit is functionally required for the proliferation of hemopoietic progenitor cells at various differentiation stages in adult bone marrow. However, the absence of functional SLF and c-Kit in fetuses with mutant alleles of Sl and W loci produces only minor effects on the myeloid and early erythroid progenitor cells in the fetal liver, although the level of the late erythroid progenitor cells is significantly affected. We used an anti-c-Kit monoclonal antibody to investigate the expression and function of c-Kit in murine fetal hemopoietic progenitor cells. Flow-cytometric analysis showed that hemopoiesis in the yolk sac and fetal liver started from cells that express c-Kit. The c-Kit expression decreased upon maturation into erythrocytes in each organ. By fluorescence activated cell sorting, the c-Kit+ cell population was enriched with the hemopoietic progenitor cells clonable in vitro (CFU-E, BFU-E and GM-CFC). To elucidate whether c-Kit functions in these progenitor cells in vivo, we took advantage of the antagonistic anti-c-Kit monoclonal antibody, ACK2, which can block the function of c-Kit. Administration of ACK2 after 12.5 days of gestation rapidly eliminated BFU-E and GM-CFC as well as CFU-E from the fetal liver. However, the number of these progenitor cells in the yolk sac and fetal liver was less affected when the fetuses were given ACK2 before 12.5 days of gestation. Our results provide evidence that there are two waves of hemopoiesis in murine embryos relative to c-Kit dependency. The c-Kit has an essential role on the growth of hemopoietic progenitor cells in the fetal liver after 12.5 days of gestation, whereas the progenitor cells in the liver and yolk sac of the earlier embryo do not depend on c-Kit and its ligand SLF.

scholarly journals Isolation of murine fetal hemopoietic progenitor cells and selective fractionation of various erythroid precursors

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 376-386 ◽  
Author(s):  
NA Nicola ◽  
D Metcalf ◽  
H von Melchner ◽  
AW Burgess
Keyword(s):  
Progenitor Cells ◽  
Fetal Liver ◽  
Pokeweed Mitogen ◽  
Erythroid Progenitor ◽  
Clonal Proliferation ◽  
Erythroid Progenitor Cells ◽  
Blast Cells ◽  
Hemopoietic Progenitor ◽  
Hemopoietic Progenitor Cells

Abstract Hemopoietic progenitor cells (colony- and cluster-forming cells in semisolid agar) were purified from light density CBA murine fetal liver cells using fluorescein-conjugated pokeweed mitogen (PWM) and a rhodamine-conjugated antineutrophil serum sandwich (alpha N) and three- parameter fluorescence-activated cell sorting. All clonable progenitor cells were highly enriched (36–50-fold) in PWM-positive (greater than channel 15), alpha N-negative (less than channel 30) fractions with relatively high intensity (greater than 100) low angle light scatter. No separation was achieved between different types of progenitor cells (granulocyte-macrophage and erythroid colony-forming cells). The enriched fraction was a pure population of large, basophilic, undifferentiated blast cells, and in agar cultures stimulated with colony-stimulating factors, up to 90% of the enriched cells were hemopoietic progenitor cells capable of varying levels of clonal proliferation. Further fractionation based on increasing fluorescence with PWM separated into discrete populations, nonproliferative morphologically recognizable erythroid cells, late erythroid progenitor cells (day 2 CFU-E), and cells forming pure or mixed erythroid burst colonies. In addition, the majority of pluripotential hemopoietic stem cells (CFU-SS) were clearly separated from progenitor cells forming colonies in vitro. The present techniques provide suitable numbers of enriched progenitor cells for a variety of biological and biochemical studies.

Spatial localization of transplanted hemopoietic stem cells: inferences for the localization of stem cell niches

Blood ◽  
2001 ◽  
Vol 97 (8) ◽  
pp. 2293-2299 ◽  
Author(s):  
Susan K. Nilsson ◽  
Hayley M. Johnston ◽  
Judi A. Coverdale
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Spatial Distribution ◽  
Stem Cell ◽  
Progenitor Cells ◽  
Hemopoietic Stem Cell ◽  
In Situ Fixation ◽  
Stem Cell Niches ◽  
Hemopoietic Progenitor ◽  
Hemopoietic Progenitor Cells

Abstract The spatial distribution of subpopulations of hemopoietic progenitor cells following syngeneic transplantation was investigated at the single-cell level. The location of infused hemopoietic progenitor cells within the femoral bone marrow of nonablated recipients was determined by 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester labeling of cells and in situ fixation by perfusion. Analysis performed over 15 hours after infusion demonstrated that the spatial distribution of transplanted marrow cells is not a random process. Although the majority of cells enter the bone marrow from the central marrow vessels, the subsequent localization within the bone marrow varied according to their phenotype. Candidate “stem cells” demonstrated selective redistribution and were significantly enriched within the endosteal region, whereas mature terminally differentiated and lineage-committed cells selectively redistributed away from the endosteal region and were predominantly in the central marrow region. Together, these data strongly support historical evidence of the presence of endosteal hemopoietic stem cell niches.

scholarly journals Isolation of murine fetal hemopoietic progenitor cells and selective fractionation of various erythroid precursors

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 376-386
Author(s):  
NA Nicola ◽  
D Metcalf ◽  
H von Melchner ◽  
AW Burgess
Keyword(s):  
Progenitor Cells ◽  
Fetal Liver ◽  
Pokeweed Mitogen ◽  
Erythroid Progenitor ◽  
Clonal Proliferation ◽  
Erythroid Progenitor Cells ◽  
Blast Cells ◽  
Hemopoietic Progenitor ◽  
Hemopoietic Progenitor Cells

Hemopoietic progenitor cells (colony- and cluster-forming cells in semisolid agar) were purified from light density CBA murine fetal liver cells using fluorescein-conjugated pokeweed mitogen (PWM) and a rhodamine-conjugated antineutrophil serum sandwich (alpha N) and three- parameter fluorescence-activated cell sorting. All clonable progenitor cells were highly enriched (36–50-fold) in PWM-positive (greater than channel 15), alpha N-negative (less than channel 30) fractions with relatively high intensity (greater than 100) low angle light scatter. No separation was achieved between different types of progenitor cells (granulocyte-macrophage and erythroid colony-forming cells). The enriched fraction was a pure population of large, basophilic, undifferentiated blast cells, and in agar cultures stimulated with colony-stimulating factors, up to 90% of the enriched cells were hemopoietic progenitor cells capable of varying levels of clonal proliferation. Further fractionation based on increasing fluorescence with PWM separated into discrete populations, nonproliferative morphologically recognizable erythroid cells, late erythroid progenitor cells (day 2 CFU-E), and cells forming pure or mixed erythroid burst colonies. In addition, the majority of pluripotential hemopoietic stem cells (CFU-SS) were clearly separated from progenitor cells forming colonies in vitro. The present techniques provide suitable numbers of enriched progenitor cells for a variety of biological and biochemical studies.

Sign in / Sign up

Export Citation Format

Share Document