ABSTRACT
A pregnant mouse uterus and embryo extract (PMUE) that contains growth hematopoietic factor (M-CSF or CSF-1), was used to test its action on the phagocytic and digestive functions of macrophage. Macrophages incubated with and without PMUE for 24 hours previous to each experiment were compared. A good phagocytosis of Trypanosoma cruzi by macrophages incubated with PMUE, was observed on video microscopy. No phagocytic activity was observed in the macrophages deprived of PMUE 24 hours before. The studies of phagocytic and degradative behavior of macrophages by both soluble and particulated (S. aureus) complex 125I-antibodies showed that total binding of soluble ligands was almost double in the group of macrophages incubated with PMUE. Both the soluble and particulated ligands were digested more efficiently by the macrophages stimulated by PMUE. Counting the macrophages with trypan blue, an equal viability was found, of the cells incubated with and without PMUE. From the experimental data obtained, we may conclude that the hematopoietic growth factor present in PMUE is essential for phagocytic and degradative functions of macrophages.
Subject(s)
Macrophage Activation/drug effects , Macrophage Colony-Stimulating Factor/pharmacology , Animals , Embryo, Mammalian/chemistry , Embryo, Nonmammalian , Female , In Vitro Techniques , Macrophage Activation/physiology , Macrophage Colony-Stimulating Factor/isolation & purification , Mice , Phagocytosis/drug effects , Pregnancy , Trypanosoma cruzi , Uterus/chemistryABSTRACT
A pregnant mouse uterus and embryo extract (PMUE) that contains growth hematopoietic factor (M-CSF or CSF-1), was used to test its action on the phagocytic and digestive functions of macrophage. Macrophages incubated with and without PMUE for 24 hours previous to each experiment were compared. A good phagocytosis of Trypanosoma cruzi by macrophages incubated with PMUE, was observed on video microscopy. No phagocytic activity was observed in the macrophages deprived of PMUE 24 hours before. The studies of phagocytic and degradative behavior of macrophages by both soluble and particulated (S. aureus) complex 125I-antibodies showed that total binding of soluble ligands was almost double in the group of macrophages incubated with PMUE. Both the soluble and particulated ligands were digested more efficiently by the macrophages stimulated by PMUE. Counting the macrophages with trypan blue, an equal viability was found, of the cells incubated with and without PMUE. From the experimental data obtained, we may conclude that the hematopoietic growth factor present in PMUE is essential for phagocytic and degradative functions of macrophages.(AU)
Subject(s)
Animals , Female , Pregnancy , Mice , In Vitro Techniques , RESEARCH SUPPORT, NON-U.S. GOVT , Macrophage Activation/drug effects , Macrophage Colony-Stimulating Factor/pharmacology , Embryonic Structures/chemistry , Macrophage Activation/physiology , Macrophage Colony-Stimulating Factor/isolation & purification , Phagocytosis/drug effects , Trypanosoma cruzi , Uterus/chemistryABSTRACT
A pregnant mouse uterus and embryo extract (PMUE) that contains growth hematopoietic factor (M-CSF or CSF-1), was used to test its action on the phagocytic and digestive functions of macrophage. Macrophages incubated with and without PMUE for 24 hours previous to each experiment were compared. A good phagocytosis of Trypanosoma cruzi by macrophages incubated with PMUE, was observed on video microscopy. No phagocytic activity was observed in the macrophages deprived of PMUE 24 hours before. The studies of phagocytic and degradative behavior of macrophages by both soluble and particulated (S. aureus) complex 125I-antibodies showed that total binding of soluble ligands was almost double in the group of macrophages incubated with PMUE. Both the soluble and particulated ligands were digested more efficiently by the macrophages stimulated by PMUE. Counting the macrophages with trypan blue, an equal viability was found, of the cells incubated with and without PMUE. From the experimental data obtained, we may conclude that the hematopoietic growth factor present in PMUE is essential for phagocytic and degradative functions of macrophages.
Subject(s)
Animals , Female , Pregnancy , Mice , Macrophage Activation/drug effects , Macrophage Colony-Stimulating Factor/pharmacology , In Vitro Techniques , Macrophage Activation/physiology , Embryonic Structures/chemistry , Macrophage Colony-Stimulating Factor/isolation & purification , Phagocytosis , Trypanosoma cruzi , Uterus/chemistryABSTRACT
This study concerns the role of endogenous polyamines in the proliferation of normal hematopoietic progenitor cells: high-proliferative potential colony-forming cells (HPP-CFC), and low-proliferative potential colony-forming cells (LPP-CFC) in CFW/ep mouse bone marrow cells in the agar culture system. DL-a-difluoromethylornithine (DFMO) was used as a selective and irreversible inhibitor of ornithine decarboxylase which is the first limiting step in polyamine biosynthesis. The polyamines have been implicated in cell proliferation and differentiation. The results showed that DFMO significantly inhibited the formation of LPP-CFC colonies and completely inhibited the growth of the HPP-CFC colonies. Addition of exogenous putrescine at the concentration of 10(-7) M reverses the suppressive action of DFMO in both progenitor cells. At this concentration putrescine alone had no affect on the number or the size of the colonies. It was then concluded that endogenous polyamines appear to be essential for HPP-CFC proliferation and an important requirement for LPP-CFC proliferation.
Subject(s)
Hematopoietic Stem Cells/cytology , Polyamines/pharmacology , Animals , Antineoplastic Agents/pharmacology , Bone Marrow Cells , Cell Division/drug effects , Cells, Cultured/cytology , Cells, Cultured/drug effects , Eflornithine/pharmacology , Female , Hematopoietic Stem Cells/drug effects , Mice , Mice, Inbred Strains , Putrescine/pharmacologyABSTRACT
This study concerns the role of endogenous polyamines in the proliferation of normal hematopoietic progenitor cells: high-proliferative potential colony-forming cells (HPP-CFC), and low-proliferative potential colony-forming cells (LPP-CFC) in CFW/ep mouse bone marrow cells in the agar culture system. DL-a-difluoromethylornithine (DFMO) was used as a selective and irreversible inhibitor of ornithine decarboxylase which is the first limiting step in polyamine biosynthesis. The polyamines have been implicated in cell proliferation and differentiation. The results showed that DFMO significantly inhibited the formation of LPP-CFC colonies and completely inhibited the growth of the HPP-CFC colonies. Addition of exogenous putrescine at the concentration of 10(-7) M reverses the suppressive action of DFMO in both progenitor cells. At this concentration putrescine alone had no affect on the number or the size of the colonies. It was then concluded that endogenous polyamines appear to be essential for HPP-CFC proliferation and an important requirement for LPP-CFC proliferation.(AU)
Subject(s)
Animals , Female , Mice , RESEARCH SUPPORT, NON-U.S. GOVT , Hematopoietic Stem Cells/cytology , Polyamines/pharmacology , Antineoplastic Agents/pharmacology , Bone Marrow Cells , Cell Division/drug effects , Cells, Cultured/cytology , Cells, Cultured/drug effects , Eflornithine/pharmacology , Hematopoietic Stem Cells/drug effects , Mice, Inbred Strains , Putrescine/pharmacologyABSTRACT
This study concerns the role of endogenous polyamines in the proliferation of normal hematopoietic progenitor cells: high-proliferative potential colony-forming cells (HPP-CFC), and low-proliferative potential colony-forming cells (LPP-CFC) in CFW/ep mouse bone marrow cells in the agar culture system. DL-a-difluoromethylornithine (DFMO) was used as a selective and irreversible inhibitor of ornithine decarboxylase which is the first limiting step in polyamine biosynthesis. The polyamines have been implicated in cell proliferation and differentiation. The results showed that DFMO significantly inhibited the formation of LPP-CFC colonies and completely inhibited the growth of the HPP-CFC colonies. Addition of exogenous putrescine at the concentration of 10(-7) M reverses the suppressive action of DFMO in both progenitor cells. At this concentration putrescine alone had no affect on the number or the size of the colonies. It was then concluded that endogenous polyamines appear to be essential for HPP-CFC proliferation and an important requirement for LPP-CFC proliferation.
Subject(s)
Animals , Female , Mice , Hematopoietic Stem Cells/cytology , Polyamines , Antineoplastic Agents/pharmacology , Bone Marrow Cells , Mice, Inbred Strains , Cells, Cultured/cytology , Cells, Cultured/drug effects , Hematopoietic Stem Cells/drug effects , Cell Division/drug effects , Eflornithine , PutrescineABSTRACT
The interaction of macrophages and T. cruzi has been studied in vitro culture under three different temperatures. After 24 hours incubation at 29 degree C a large number of recognizable parasites inside macrophages is observed with evidences of cell divisions. At video microscopy they show a slow motion and a predominance of epimastigotes and some round shapes (amastigotes). This was corroborated at the light and electron microscopes. No evidence of lysis in the phagosome vacuoles was observed. At 40 degrees C, macrophages show a large number of residual bodies and phagocytic vacuoles with digested parasites. At 37 degrees C, an intermediate stage with normal and digested parasites inside macrophages is observed. No significant evidences of affected phagocytic and degradative properties of the macrophages were obtained at those temperatures. It is postulated that temperature mainly affects the parasite resistance to intracellular digestion.
Subject(s)
Macrophages/parasitology , Trypanosoma cruzi/physiology , Animals , Cells, Cultured , Macrophages/ultrastructure , Mice , Microscopy, Electron , Phagocytosis , Photomicrography , Temperature , Trypanosoma cruzi/ultrastructure , Vacuoles/parasitology , Video RecordingABSTRACT
Se estudió la interacción de macrófagos y T.cruzi cultivados in vitro a tres diferentes temperaturas. Luego de 24 horas de incubación a 29-C se observó un gran número de parásitos dentro de los macrófagos con evidencias de división celular. Estos parásitos presentaban un predominio de formas epimastihotas y algunas redondeadas (amastigotes) y movimientos lentos vistos por videomicroscopía. Se corroboró esta observación con los microscopios de luz y electrónico. No se observó evidencias de lisis en los fagosomas. A 40-C los macrófagos muestran un gran número de cuerpos residuales y vacuolas fagocíticas con parásitos digeridos. A 37-C se observó un estado intermedio con parásitos digeridos y normales. Se comprobó que estas temperaturas no afectan al macrófago en su capacidad fagocítica y digestiva. Se postula que la temperatura afecta principalmente al parásito en su resistencia a la digestión intracelular
Subject(s)
Animals , Mice , Macrophages/parasitology , Trypanosoma cruzi/physiology , Cells, Cultured , Macrophages/ultrastructure , Microscopy, Electron , Phagocytes , Photomicrography , Temperature , Trypanosoma cruzi/ultrastructure , Vacuoles/parasitology , Video RecordingABSTRACT
Se estudió la interacción de macrófagos y T.cruzi cultivados in vitro a tres diferentes temperaturas. Luego de 24 horas de incubación a 29-C se observó un gran número de parásitos dentro de los macrófagos con evidencias de división celular. Estos parásitos presentaban un predominio de formas epimastihotas y algunas redondeadas (amastigotes) y movimientos lentos vistos por videomicroscopía. Se corroboró esta observación con los microscopios de luz y electrónico. No se observó evidencias de lisis en los fagosomas. A 40-C los macrófagos muestran un gran número de cuerpos residuales y vacuolas fagocíticas con parásitos digeridos. A 37-C se observó un estado intermedio con parásitos digeridos y normales. Se comprobó que estas temperaturas no afectan al macrófago en su capacidad fagocítica y digestiva. Se postula que la temperatura afecta principalmente al parásito en su resistencia a la digestión intracelular (AU)
Subject(s)
Animals , Mice , Macrophages/parasitology , Trypanosoma cruzi/physiology , Cells, Cultured , Macrophages/ultrastructure , Microscopy, Electron , Phagocytes , Photomicrography , Temperature , Trypanosoma cruzi/ultrastructure , Vacuoles/parasitology , Video RecordingABSTRACT
The interaction of macrophages and T. cruzi has been studied in vitro culture under three different temperatures. After 24 hours incubation at 29 degree C a large number of recognizable parasites inside macrophages is observed with evidences of cell divisions. At video microscopy they show a slow motion and a predominance of epimastigotes and some round shapes (amastigotes). This was corroborated at the light and electron microscopes. No evidence of lysis in the phagosome vacuoles was observed. At 40 degrees C, macrophages show a large number of residual bodies and phagocytic vacuoles with digested parasites. At 37 degrees C, an intermediate stage with normal and digested parasites inside macrophages is observed. No significant evidences of affected phagocytic and degradative properties of the macrophages were obtained at those temperatures. It is postulated that temperature mainly affects the parasite resistance to intracellular digestion.
ABSTRACT
In this paper we describe a crude pregnant mouse uterus and embryo extract (PMUE) prepared from CFW/ep mice which was able to stimulate the proliferation of high-proliferative-potential colony-forming cells (HPP-CFC) of bone marrow of normal mice, in vitro, in semisolid agar culture system. The development of that primitive murine progenitor cells requires the presence of a macrophage-stimulating factor (CSF-1) plus a synergistic factor (SF). The biological activity of both factors was present in our extracts. The higher SF activity was found in uterine plus placental tissues extracts. The SF was precipitated over 45 per cent ammonium sulfate saturation, and behaved as a nondialyzable substance, remained unaffected by trypsin digestion, and was heat-stable (70 degrees C for 15 min).
Subject(s)
Bone Marrow Cells , Colony-Stimulating Factors/analysis , Embryo, Mammalian/analysis , Uterus/analysis , Animals , Colony-Stimulating Factors/isolation & purification , Colony-Stimulating Factors/pharmacology , Female , Growth Substances/isolation & purification , Growth Substances/pharmacology , Hematopoietic Cell Growth Factors , In Vitro Techniques , Macrophage Colony-Stimulating Factor , Mice , Mice, Inbred Strains , Pregnancy , Trypsin/metabolismABSTRACT
In this paper we describe a crude pregnant mouse uterus and embryo extract (PMUE) prepared from CFW/ep mice which was able to stimulate the proliferation of high-proliferative-potential colony-forming cells (HPP-CFC) of bone marrow of normal mice, in vitro, in semisolid agar culture system. The development of that primitive murine progenitor cells requires the presence of a macrophage-stimulating factor (CSF-1) plus a synergistic factor (SF). The biological activity of both factors was present in our extracts. The higher SF activity was found in uterine plus placental tissues extracts. The SF was precipitated over 45 per cent ammonium sulfate saturation, and behaved as a nondialyzable substance, remained unaffected by trypsin digestion, and was heat-stable (70 degrees C for 15 min).
ABSTRACT
Se estudió la ación "in vitro" del extracto de embriones y úteros de ratón preñado (PMUE), que promueve el desarrollo de grandes colonias de células progenitoras comprometidas de alto potencial prolifarativo (HPP-CFC) y su diferenciación a macrófagos. El extracto se obtuvo de la cepa CFW/ep y tanto las células de la médula ósea de ésta como las de ratones suizos normales se cultivaron en medio semisólido con agar para demostrar la acción biológica del PMUE y medio condicionante de placenta humana (HPCM). La proliferación de esas células progenitoreas muy primitivas requiere la presencia concurrente de un factor estimulante de macrófagos - CSF - y de un factor sinergístico (SF). La actividad biológica de ambos está presente en el extracto, encontrándose la mayor actividad en útero y placenta. El SF precipita con sulfato de amonio con más de 45% de saturación, no es dializable y es termiestable (a 70-C durante 15 min). Se destaca su resistencia a la acción enzimática de la tripsina (AU)
Subject(s)
Pregnancy , Mice , Animals , Female , In Vitro Techniques , Stem Cells/physiology , Uterus/analysis , Embryonic Structures/analysis , Bone Marrow/cytology , Colony-Stimulating Factors/pharmacology , Trypsin/metabolism , Colony-Stimulating Factors/isolation & purification , Mice, Inbred StrainsABSTRACT
Se estudió la ación "in vitro" del extracto de embriones y úteros de ratón preñado (PMUE), que promueve el desarrollo de grandes colonias de células progenitoras comprometidas de alto potencial prolifarativo (HPP-CFC) y su diferenciación a macrófagos. El extracto se obtuvo de la cepa CFW/ep y tanto las células de la médula ósea de ésta como las de ratones suizos normales se cultivaron en medio semisólido con agar para demostrar la acción biológica del PMUE y medio condicionante de placenta humana (HPCM). La proliferación de esas células progenitoreas muy primitivas requiere la presencia concurrente de un factor estimulante de macrófagos - CSF - y de un factor sinergístico (SF). La actividad biológica de ambos está presente en el extracto, encontrándose la mayor actividad en útero y placenta. El SF precipita con sulfato de amonio con más de 45% de saturación, no es dializable y es termiestable (a 70-C durante 15 min). Se destaca su resistencia a la acción enzimática de la tripsina