scholarly journals Characteristics of murine megakaryocytic colonies in vitro

Blood ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 169-179 ◽  
Author(s):  
SA Burstein ◽  
JW Adamson ◽  
D Thorning ◽  
LA Harker
Keyword(s):  
Conditioned Medium ◽  
Suicide Rate ◽  
Tritiated Thymidine ◽  
Colony Growth ◽  
Velocity Sedimentation ◽  
Agar Gel ◽  
Colony Forming Units ◽  
Granulocytic Colony

Abstract Characteristics of murine megakaryocytic colonies and their progenitor cells (CFU-m) were studied in vitro in agar gel. Colony growth required the presence of poke-weed-mitogen-stimulated spleen-conditioned medium. The number of colonies formed was linearly related to both the number of marrow cells plated and the amount of conditioned medium added. In addition, CFU-m were found in both the spleen and peripheral blood. Conditioned medium was also made without plasma, and this resulted in a cloning efficiency greater than that of conditioned medium prepared with plasma. The percentage of CFU-m in DNA synthesis was low (10%), as determined both in vivo and in vitro. Velocity sedimentation revealed that the majority of CFU-m sedimented at 4.3 mm/hr and had a tritiated thymidine (3H-TdR) suicide rate of 1.5 +/- 1.5%. A shoulder on the profile of CFU-m sedimented at approximately 6 mm/hr, with a suicide rate of 79 +/- 2%. Analysis of these data indicated that the majority of CFU-m were not in cycle or were in a long G1 period. The results suggest that CFU-m is a primitive progenitor, possibly closely related to murine splenic colony-forming units (CFU-s), analogous to erythroid bursts and granulocytic colony-forming units.

scholarly journals Characteristics of murine megakaryocytic colonies in vitro

Blood ◽  
1979 ◽  
Vol 54 (1) ◽  
pp. 169-179 ◽  
Author(s):  
SA Burstein ◽  
JW Adamson ◽  
D Thorning ◽  
LA Harker
Keyword(s):  
Conditioned Medium ◽  
Suicide Rate ◽  
Tritiated Thymidine ◽  
Colony Growth ◽  
Velocity Sedimentation ◽  
Agar Gel ◽  
Colony Forming Units ◽  
Granulocytic Colony

Characteristics of murine megakaryocytic colonies and their progenitor cells (CFU-m) were studied in vitro in agar gel. Colony growth required the presence of poke-weed-mitogen-stimulated spleen-conditioned medium. The number of colonies formed was linearly related to both the number of marrow cells plated and the amount of conditioned medium added. In addition, CFU-m were found in both the spleen and peripheral blood. Conditioned medium was also made without plasma, and this resulted in a cloning efficiency greater than that of conditioned medium prepared with plasma. The percentage of CFU-m in DNA synthesis was low (10%), as determined both in vivo and in vitro. Velocity sedimentation revealed that the majority of CFU-m sedimented at 4.3 mm/hr and had a tritiated thymidine (3H-TdR) suicide rate of 1.5 +/- 1.5%. A shoulder on the profile of CFU-m sedimented at approximately 6 mm/hr, with a suicide rate of 79 +/- 2%. Analysis of these data indicated that the majority of CFU-m were not in cycle or were in a long G1 period. The results suggest that CFU-m is a primitive progenitor, possibly closely related to murine splenic colony-forming units (CFU-s), analogous to erythroid bursts and granulocytic colony-forming units.

scholarly journals Human myeloma in vitro colony growth: interrelationships between drug sensitivity, cell kinetics, and patient survival duration

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 929-934 ◽  
Author(s):  
BG Durie ◽  
LA Young ◽  
SE Salmon
Keyword(s):  
Drug Sensitivity ◽  
Cell Kinetics ◽  
Plasma Cells ◽  
Tritiated Thymidine ◽  
Colony Growth ◽  
Survival Duration ◽  
Sensitivity Testing ◽  
Selective Sensitivity

Abstract Ninety-seven patients with multiple myeloma evaluated serially had both a tritiated thymidine labeling index of bone marrow plasma cells (LI%) and in vitro myeloma stem cell culture performed. Thirty-three patients with myeloma colony growth had in vitro drug sensitivity testing carried out, 18 having in addition in vitro thymidine suicide determinations. The LI% and the likelihood of in vitro myeloma colony growth were highly correlated: the higher the LI%, the more likely was colony or cluster growth (p less than 0.001). The tritiated thymidine suicide of myeloma stem cells was usually very high. There was excellent correlation between in vitro and in vivo drug sensitivity. Both pretreatment drug resistance and selective sensitivity (e.g., interferon, bisantrene, methotrexate, vinblastine) at the time of relapse were accurately detected and correlated well with survival duration (p = 0.01 Wilcoxan). Although LI% and in vitro sensitivity were clearly independent variables, a high LI% (greater than 3%) plus in vitro resistance were associated with a subsequent survival duration of less than 6 mo. The studies allowed dissection of the complex interrelationship between cell kinetics and drug sensitivity.

scholarly journals Steroids and hematopoiesis. III. The response of granulocytic and erythroid colony-forming cells to steroids of different classes

Blood ◽  
1976 ◽  
Vol 48 (6) ◽  
pp. 855-864 ◽  
Author(s):  
JW Singer ◽  
JW Adamson
Keyword(s):  
Cell Cycle ◽  
Colony Growth ◽  
Target Cells ◽  
Naturally Occurring ◽  
Colony Forming Units ◽  
Granulocytic Colony ◽  
In Vitro Response ◽  
Stimulating Factor ◽  
Do So

Selected androgenic and nonandrogenic steroids enhance in vitro granulocytic and erythroid colony formation by mouse marrow cells, but do so by influencing either different target cells or cells in different states of cell cycle. Etiocholanolone, a naturally occurring nonandrogenic testosterone metabolite, permits cells not in active cycle to respond to colony-stimulating factor or erythropoietin. Fluoxymesterone, a synthetic androgen, appears to enhance colony growth by increasing the responsiveness of target cells to tropic stimuli. The majority of cells responding to this androgen are in active DNA synthesis. Direct comparison, however, of etiocholanolone-dependent erythroid or granulocytic colony-forming cells demonstrates nonidentity of the target cells. Thus colony-forming units responding to different classes of steroids are in different states of cell cycle and are physically separable. The enhancement of the in vitro response of colony-forming cells to regulating hormones by steroids such as etiocholanolane suggests a mechanism by which such agents may be therapeutically effective in certain cases of marrow failure in man.

scholarly journals Human myeloma in vitro colony growth: interrelationships between drug sensitivity, cell kinetics, and patient survival duration

Blood ◽  
1983 ◽  
Vol 61 (5) ◽  
pp. 929-934
Author(s):  
BG Durie ◽  
LA Young ◽  
SE Salmon
Keyword(s):  
Drug Sensitivity ◽  
Cell Kinetics ◽  
Plasma Cells ◽  
Tritiated Thymidine ◽  
Colony Growth ◽  
Survival Duration ◽  
Sensitivity Testing ◽  
Selective Sensitivity

Ninety-seven patients with multiple myeloma evaluated serially had both a tritiated thymidine labeling index of bone marrow plasma cells (LI%) and in vitro myeloma stem cell culture performed. Thirty-three patients with myeloma colony growth had in vitro drug sensitivity testing carried out, 18 having in addition in vitro thymidine suicide determinations. The LI% and the likelihood of in vitro myeloma colony growth were highly correlated: the higher the LI%, the more likely was colony or cluster growth (p less than 0.001). The tritiated thymidine suicide of myeloma stem cells was usually very high. There was excellent correlation between in vitro and in vivo drug sensitivity. Both pretreatment drug resistance and selective sensitivity (e.g., interferon, bisantrene, methotrexate, vinblastine) at the time of relapse were accurately detected and correlated well with survival duration (p = 0.01 Wilcoxan). Although LI% and in vitro sensitivity were clearly independent variables, a high LI% (greater than 3%) plus in vitro resistance were associated with a subsequent survival duration of less than 6 mo. The studies allowed dissection of the complex interrelationship between cell kinetics and drug sensitivity.

scholarly journals Steroids and hematopoiesis. III. The response of granulocytic and erythroid colony-forming cells to steroids of different classes

Blood ◽  
1976 ◽  
Vol 48 (6) ◽  
pp. 855-864 ◽  
Author(s):  
JW Singer ◽  
JW Adamson
Keyword(s):  
Cell Cycle ◽  
Colony Growth ◽  
Target Cells ◽  
Naturally Occurring ◽  
Colony Forming Units ◽  
Granulocytic Colony ◽  
In Vitro Response ◽  
Stimulating Factor ◽  
Do So

Abstract Selected androgenic and nonandrogenic steroids enhance in vitro granulocytic and erythroid colony formation by mouse marrow cells, but do so by influencing either different target cells or cells in different states of cell cycle. Etiocholanolone, a naturally occurring nonandrogenic testosterone metabolite, permits cells not in active cycle to respond to colony-stimulating factor or erythropoietin. Fluoxymesterone, a synthetic androgen, appears to enhance colony growth by increasing the responsiveness of target cells to tropic stimuli. The majority of cells responding to this androgen are in active DNA synthesis. Direct comparison, however, of etiocholanolone-dependent erythroid or granulocytic colony-forming cells demonstrates nonidentity of the target cells. Thus colony-forming units responding to different classes of steroids are in different states of cell cycle and are physically separable. The enhancement of the in vitro response of colony-forming cells to regulating hormones by steroids such as etiocholanolane suggests a mechanism by which such agents may be therapeutically effective in certain cases of marrow failure in man.

scholarly journals Conditioned medium-preconditioned EPCs enhanced the ability in oligovascular repair in cerebral ischemia neonatal rats

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ning Zhou ◽  
Lei Wang ◽  
Ping Fu ◽  
Zihao Cui ◽  
Yuhang Ge ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Western Blot ◽  
Conditioned Medium ◽  
Cognitive Outcome ◽  
Adult Brain ◽  
Neonatal Rat ◽  
Vascular Density ◽  
Cxcr4 Axis

Abstract Background Oligovascular niche mediates interactions between cerebral endothelial cells and oligodendrocyte precursor cells (OPCs). Disruption of OPC-endothelium trophic coupling may aggravate the progress of cerebral white matter injury (WMI) because endothelial cells could not provide sufficient support under diseased conditions. Endothelial progenitor cells (EPCs) have been reported to ameliorate WMI in the adult brain by boosting oligovascular remodeling. It is necessary to clarify the role of the conditioned medium from hypoxic endothelial cells preconditioned EPCs (EC-pEPCs) in WMI since EPCs usually were recruited and play important roles under blood-brain barrier disruption. Here, we investigated the effects of EC-pEPCs on oligovascular remodeling in a neonatal rat model of WMI. Methods In vitro, OPC apoptosis induced by the conditioned medium from oxygen-glucose deprivation-injured brain microvascular endothelial cells (OGD-EC-CM) was analyzed by TUNEL and FACS. The effects of EPCs on EC damage and the expression of cytomokine C-X-C motif ligand 12 (CXCL12) were examined by western blot and FACS. The effect of the CM from EC-pEPCs against OPC apoptosis was also verified by western blot and silencing RNA. In vivo, P3 rat pups were subjected to right common carotid artery ligation and hypoxia and treated with EPCs or EC-pEPCs at P7, and then angiogenesis and myelination together with cognitive outcome were evaluated at the 6th week. Results In vitro, EPCs enhanced endothelial function and decreased OPC apoptosis. Meanwhile, it was confirmed that OGD-EC-CM induced an increase of CXCL12 in EPCs, and CXCL12-CXCR4 axis is a key signaling since CXCR4 knockdown alleviated the anti-apoptosis effect of EPCs on OPCs. In vivo, the number of EPCs and CXCL12 protein level markedly increased in the WMI rats. Compared to the EPCs, EC-pEPCs significantly decreased OPC apoptosis, increased vascular density and myelination in the corpus callosum, and improved learning and memory deficits in the neonatal rat WMI model. Conclusions EC-pEPCs more effectively promote oligovascular remodeling and myelination via CXCL12-CXCR4 axis in the neonatal rat WMI model.

Separation By Velocity Sedimentation of Human Haemopoietic Precursors Forming Colonies in vivo and in vitro Cultures

1985 ◽  
Vol 18 (4) ◽  
pp. 399-406
Author(s):  
E. Niskanen ◽  
J. R. Wells ◽  
D. W. Golde ◽  
M. J. Cline
Keyword(s):  
In Vitro Cultures ◽  
Velocity Sedimentation

scholarly journals Fitness of Isolates of Phytophthora capsici Resistant to Mefenoxam from Squash and Pepper Fields in North Carolina

Plant Disease ◽  
2008 ◽  
Vol 92 (10) ◽  
pp. 1439-1443 ◽  
Author(s):  
Adalberto C. Café-Filho ◽  
Jean Beagle Ristaino
Keyword(s):  
North Carolina ◽  
Phytophthora Capsici ◽  
Colony Growth ◽  
Relative Fitness ◽  
In Planta ◽  
Phytophthora Blight ◽  
In Vivo Experiments ◽  
First Time

Despite the wide adoption of mefenoxam (Ridomil Gold EC) for vegetables in North Carolina, the incidence of Phytophthora blight on pepper (Capsicum annuum) and squash (Cucurbita pepo) is high. Seventy-five isolates of Phytophthora capsici were collected in five pepper and one squash field in order to assess mefenoxam sensitivity. The relative fitness of resistant and sensitive isolates was contrasted in vitro by their respective rates of colony growth and their ability to produce sporangia in unamended V8 juice agar medium. In in vivo experiments, the aggressiveness of isolates on pepper was evaluated. The frequency of resistant isolates in North Carolina populations was 63%, considerably higher than resistance levels in areas where mefenoxam is not widely adopted. Resistant isolates grew on amended media at rates >80 to 90% and >100% of the nonamended control at 100 μg ml-1 and 5 μg ml-1, respectively. Sensitive isolates did not growth at 5 or 100 μg ml-1. All isolates from three fields, including two pepper and a squash field, were resistant to mefenoxam. Populations from other fields were composed of either mixes of sensitive and resistant isolates or only sensitive isolates. Response to mefenoxam remained stable during the course of in vitro and in planta experiments. Occurrence of a mefenoxam-resistant population of P. capsici on squash is reported here for the first time in North Carolina. When measured by rate of colony growth, sporulation in vitro, or aggressiveness in planta, fitness of resistant isolates was not reduced. Mefenoxam-resistant isolates from squash were as aggressive on pepper as sensitive or resistant pepper isolates. These results suggest that mefenoxam-resistant populations of P. capsici are as virulent and fit as sensitive populations.

scholarly journals 1222. Are Reduced Concentrations of Chlorine-Based Disinfectants Effective Against Candida auris?

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S439-S439
Author(s):  
Jessica Kumar ◽  
Jennifer Cadnum ◽  
Y Karen Ng Wong ◽  
Thriveen Sankar Chittoor Mana ◽  
Heba Alhmidi ◽  
...  
Keyword(s):  
Sodium Hypochlorite ◽  
Exposure Time ◽  
Environmental Protection Agency ◽  
Simulated Patient ◽  
Patient Room ◽  
Candida Auris ◽  
Colony Forming Units ◽  
Sodium Dichloroisocyanurate

Abstract Background Currently, sporicidal disinfectants such as bleach are recommended for daily and terminal disinfection of the rooms of patients with Candida auris colonization and/or infection. However, bleach and other chlorine-based disinfectants can have adverse effects on surfaces and personnel. Disinfectant solutions with reduced chlorine concentrations are commonly used for other pathogens, but it is not known if diluted or alternative products maintain efficacy against C. auris both in vitro and in vivo. Methods We tested the efficacy of different concentrations of a sodium dichloroisocyanurate (NaDCC) product and sodium hypochlorite using the method recommended by the Environmental Protection Agency (EPA) for evaluation of the efficacy of liquid disinfectants against C. auris (EPA MLB SOP MB-35-00) and in a simulated patient room. Carriers were exposed to each disinfectant for 1 and 2 minutes. Log reductions were calculated by subtracting viable organisms recovered after disinfectant exposure vs. deionized water controls. Results As shown in the figure, the NaDCC product at 4306 ppm tested with a 2 minute contact time reduced C. auris by ≥5 log10 colony-forming units (CFU) but had reduced efficacy with shorter exposure time or lower concentrations. Sodium hypochlorite was effective with 1 or 2 minute exposure times at a concentration of 6,500 ppm, and was effective at 4,000 ppm with an exposure time of 2 minutes. In the simulated patient room, NaDCC reduced C. auris contamination by ≥6 log10 CFUs on all surfaces. Conclusion A chlorine-based NaDCC product was effective at reducing C. auris. Both NaDCC and sodium hypochlorite products exhibited reduced efficacy at lower concentrations, particularly at concentrations below 4000 ppm. The NaDCC products were also effective in reducing contamination in the simulated patient room. UV-C treatment was an effective adjunct to manual cleaning. Disclosures All authors: No reported disclosures.

scholarly journals Cell kinetics of human tumors by in vitro bromodeoxyuridine labeling.

1989 ◽  
Vol 37 (9) ◽  
pp. 1449-1454 ◽  
Author(s):  
J S Meyer ◽  
J Nauert ◽  
S Koehm ◽  
J Hughes
Keyword(s):  
Tritiated Thymidine ◽  
S Phase ◽  
Dna Flow Cytometry ◽  
Breast Carcinomas ◽  
Brdu Labeling ◽  
The Central Nervous System ◽  
Labeling Method ◽  
Kinetics Of

We labeled active S-phase cells in primary breast carcinomas with a modified 5-bromo-2'-deoxyuridine (BrdU) procedure using a silver-enhanced colloidal gold visualization step. Separate samples of 29 tumors were labeled with BrdU or tritiated thymidine ([3H]-dThd), and the labeling indices (LI) from the two methods were equivalent (Spearman's correlation coefficient = 0.96). Three breast carcinomas were incubated in various mixes of both BrdU and [3H]-dThd and developed sequentially for each. Paired photomicrographs showed that the same nuclei were labeled by either precursor. The in vitro method yielded LIs similar to those reported after in vivo pulse BrdU labeling for tumors of the central nervous system. The BrdU LI correlated significantly (r = 0.76, p less than 0.001) with % S-phase by DNA flow cytometry in 33 breast carcinomas. The BrdU labeling method is simpler and more rapid than the [3H]-dThd procedure (1-2 days for completion for the former, 7-10 days for the latter), and it provides an equivalent measurement of proliferative index.

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