scholarly journals Intermittent blood flow in the KHT sarcoma--flow cytometry studies using Hoechst 33342

1990 ◽  
Vol 62 (2) ◽  
pp. 195-200 ◽  
Author(s):  
AI Minchinton ◽  
RE Durand ◽  
DJ Chaplin
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Hoechst 33342

The Effects of Heparin, Protamine, and Heparinase 1 on Platelets in vitro Using Whole Blood Flow Cytometry

2000 ◽  
Vol 9 (4) ◽  
pp. 808-812 ◽  
Author(s):  
Sibylle A. Kozek-Langenecker ◽  
S. Fazal Mohammad ◽  
Takahisa Masaki ◽  
Craig Kamerath ◽  
Alfred K. Cheung
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Whole Blood

461 Prostate stem cell isolation using Hoechst 33342 flow cytometry

2004 ◽  
Vol 3 (2) ◽  
pp. 118
Author(s):  
P. Gilmore ◽  
R. Bhatt ◽  
C. Hart ◽  
V. Ramani ◽  
N. George ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
Cell Isolation ◽  
Hoechst 33342 ◽  
Stem Cell Isolation

110 COMPARISON OF ANDROMED®, BIOXCELL®, AND BOTU-BOV® EXTENDERS FOR CRYOPRESERVATION OF BULL SEXED SEMEN

2007 ◽  
Vol 19 (1) ◽  
pp. 172 ◽  
Author(s):  
M. Q. Braga ◽  
R. V. R. Franco ◽  
L. F. Rodrigues ◽  
G. Galeli ◽  
K. M. Oliveira ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Semen Quality ◽  
Membrane Damage ◽  
Optical Microscope ◽  
Hoechst 33342 ◽  
Tukey Test ◽  
Progressive Motility ◽  
Significant Difference ◽  
Sexed Semen ◽  
Fort Collins

Sexing semen has become a worldwide technology now available in many countries through the use of flow cytometry for sexing mammal sperms (Johnson and Welch 1999 Theriogenology 52, 1323–1341). Because straws containing sexed semen have a low concentration, any condition that either improves or decreases freezing capabilities will considerably change semen quality. During cryopreservation, spermatozoa have been described as undergoing many changes that lead to membrane damage, which may result in decreased fertility (Watson 2000 Reprod. Fertil. Dev. 6 (Suppl 1), 481). Since many cryoprotectants are available on the market, the objective of the present study was to compare 3 different extenders for freezing sex-sorted semen. For this study, 25 ejaculates were collected from 8 bulls of different breeds, diluted, then dyed with Hoechst 33342 (Schenk et al. 1999 Theriogenology 52, 1375–1391), and sexed by flow cytometry (SX MoFlo®; DaKoCytomation, Inc., Fort Collins, CO, USA). After being cooled at 4°C for 1 h and 30 min, the sexed semen was centrifuged and diluted in AndroMed® (Minitüb, Tiefenbach, Germany), Bioxcell® (IMV, Aigle, France), or Botu-Bov® (Biotech Botucatu, Ltda., Sao Paulo, Brazil); the semen was packaged at 3 million total sperm in 0.25-mL straws and frozen in an automatic freezer (Digit cool 5300® IMV). To evaluate the freezing quality, the straws were thawed and incubated at 35°C for 15 min. The progressive motility was observed through an optical microscope (Coleman 200T). The statistical analyses were done using the SAS program (SAS Institute, Inc., Cary, NC, USA) and the Tukey test (P ≤ 0.05). Results show that there was no statistical difference between Bioxcell and AndroMed extenders (P ≤ 0.05). However, Botu-Bov extender showed a significant difference when compared with Bioxcell and AndroMed (see Table 1). It is also important to point out that 40% of the samples frozen with AndroMed showed non-aligned movement. Even though few ejaculates were used for this study, preliminary results showed that Bioxcell seemed to be the most suitable for freezing bull sexed semen. Table 1. Percentage of progressively motile spermatozoa after thawing

Comparison of Umbilical Cord Blood Derived Mononuclear Cells and Endothelial Generating Cells in Response to Ischemia in the Murine Hind-Limb Injury Model.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3973-3973
Author(s):  
Marcie R. Finney ◽  
Vincent J. Pompili ◽  
Nicholass G. Greco ◽  
Matthew J. Joseph ◽  
Daniel G. Winter ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Cord Blood ◽  
Umbilical Cord Blood ◽  
Hind Limb ◽  
Mononuclear Cells ◽  
Fold Increase ◽  
Cell Populations ◽  
Surface Phenotype ◽  
Limb Injury

Abstract Recent reports have studied the use of cell populations from bone marrow, peripheral blood and umbilical cord blood (UCB) in mediating therapeutic angiogenesis in patients with coronary artery disease. We investigated the surface phenotype of UCB derived-mononuclear cells (MNC), and endothelial generating cells (EGC) by flow cytometry and in vitro functional migration studies. We used a murine hind-limb injury ischemia model to assess in vivo efficacy of the cell populations. METHODS: MNC were isolated by density centrifugation and CD133+ cells were isolated by magnetic separation (Miltenyi). EGC were derived by adherence of CD133− cells cultured 16h on fibronectin-coated tissue culture plates in EGM2 media (Clonetics). Cell characterization included surface phenotype determined by flow cytometry for monocyte markers CD14 and CD11b, stromal markers CD73 and CD105, KDR (VEGFR2), and the receptor for SDF-1, CXCR4. Transwell plates with 5μm collagen coated filters (Costar) were used to observe chemotactic migration of MNC or EGC towards SDF-1 (100ng/mL). Following a 3 hour incubation, the cells migrated to the bottom wells was counted by flow cytometry with TruCOUNT™ tubes (BD Biosciences. NOD/SCID mice underwent right femoral artery ligation and were injected with cytokines (EGM2 media, n=10), MNC (n=7) or EGC (n=8), 1X 106 cells/mouse. Laser Doppler blood flow measurements were recorded weekly for four weeks. RESULTS: Enhanced expression of CXCR4, CD105, KDR, CD14 and CD11b was found in the EGC cells generated after 16h culture on fibronectin. Surface Phenotype of UCB MNC and EGC CD14 CD11b CD73 CD105 KDR CXCR4 MNC (n≥5) 8.1 ± 2.7 22.7 ± 6.6 5.1 ± 2.2 6.7 ± 2.1 7.1 ± 2.1 28.5 ± 5.8 EGC (n=5) 67.7 ± 9.5 80.0 ± 4.8 7.1 ± 4.0 33.8 ± 5.8 37.6 ± 6.4 64.6 ± 7.8 Functional assays showed increased migration of both MNC and EGC to SDF-1 compared to control media (4.9 ± 2.9, n=2 and 3.2 ± 0.6, n=3 fold increases respectively). With VEGF as a chemoattractant MNC exhibited a 1.5 fold increase over the negative control (n=1) and EGC showed a 1.4 ± 0.3 fold increase (n=2). In the murine hind-limb ischemia model the ratio of ischemic/non-ischemic limb blood flow was used to compare vasculogenic potential. There was improvement of blood flow 14 days after injection of the EGC cells (p=0.019). On days 21 and 28, blood flow ratio was higher than control but was not statistically significant (p=0.06). The difference between MNC and EGC was not significant at any time point (p> 0.05). Histological studies are ongoing. CONCLUSION: In summary, UCB derived EGC exhibited monocyte and stromal surface antigen expression, migrated to an SDF-1 gradient, and mediated improved vascular blood flow. Ongoing studies are focused on direct cell vs. paracrine effects underlying observed neovasculogenesis mediated by EGC.

APPLICATION OF A KINETIC WHOLE BLOOD FLOW CYTOMETRY METHOD TO STUDY FREE CALCIUM MOBILIZATION IN PLATELETS OF HYPERTENSIVE PATIENTS. EFFECT OF DOXAZOSIN

2004 ◽  
Vol 22 (Suppl. 2) ◽  
pp. S126
Author(s):  
M. Labiós Gómez ◽  
M. Martinez Silvestre ◽  
F. Gabriel Botella ◽  
V. Guiral Olivan ◽  
S. Palanca Suela ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Whole Blood ◽  
Calcium Mobilization ◽  
Free Calcium ◽  
Hypertensive Patients

EFFECT OF EPROSARTAN ON PLATELET ACTIVATION AND MICROPARTICLES FORMATION IN HYPERTENSION. STUDY BY WHOLE BLOOD FLOW CYTOMETRY

2004 ◽  
Vol 22 (Suppl. 2) ◽  
pp. S122
Author(s):  
M. Labiós Gómez ◽  
M. Martinez Silvestre ◽  
F. Gabriel Botella ◽  
V. Guiral Olivan ◽  
S. Palanca Suela ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Platelet Activation ◽  
Whole Blood
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