scholarly journals Do transplanted corneal limbal stem cells survive in vivo long term? Possible techniques to detect donor cell survival by polymerase chain reaction with the amelogenin gene and Y-specific probes

Eye ◽  
1997 ◽  
Vol 11 (6) ◽  
pp. 779-785 ◽  
Author(s):  
T R M Henderson ◽  
S H McCall ◽  
G R Taylor ◽  
B A Noble
Keyword(s):  
Stem Cells ◽  
Polymerase Chain Reaction ◽  
Cell Survival ◽  
Donor Cell ◽  
Chain Reaction ◽  
Limbal Stem Cells ◽  
Amelogenin Gene ◽  
Polymerase Chain

scholarly journals Downregulation of Caspase-2 Expression in Somitic Cells following Coculture with Chicken Notochord

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Rezgar Rahbari ◽  
Mohammad Mazani ◽  
Mohammad Ghasem Golmohammadi ◽  
Mohsen Sagha
Keyword(s):  
Polymerase Chain Reaction ◽  
Enzyme Activity ◽  
Cell Survival ◽  
Neural Tube ◽  
Chain Reaction ◽  
Survival Assay ◽  
Polymerase Chain ◽  
Caspase 2

Somites are spherical aggregations of mesodermal cells located on either sides of neural tube and are differentiated into sclerotome and dermomyotome. Notochord as an axial mesoderm has a major role in somitic cell survival and differentiation in vivo. Despite secreting the survival factors, how to notochord inhibits somitic cells apoptosis remains to be elusive. So, this study was aimed to investigate downregulation of caspase-2 expression in somitic cells upon coculturing with notochord. By using alginate system to encapsulate the isolated notochord in Somite + Notochord group, the embryonic somites were cocultured with the notochord on different days. Concurrently in somite group, the somites were cultured alone. Survival assay with MTT showed that the rate of viability in somitic cells cocultured with notochord increased from 59% on day 2 to 89.7% on day 6 but decreased to 38.5% on day 10 after coculturing. Reverse transcriptase-polymerase chain reaction and spectrophotometry analysis also confirmed these findings and showed low caspase-2 and high Bcl-2 expressions and low caspase-2 enzyme activity in somitic cells cocultured with notochord, respectively. These results clearly show that the notochord enhances survival of somitic cells in vitro through downregulating of caspase-2 expression along with triggering differentiation of somitic cells to Pax-1 expressing mesenchymal cells.

scholarly journals Maximizing Ventricular Function With Multimodal Cell-Based Gene Therapy

Circulation ◽  
2005 ◽  
Vol 112 (9_supplement) ◽  
Author(s):  
Terrence M. Yau ◽  
Christopher Kim ◽  
Guangming Li ◽  
Yaoguang Zhang ◽  
Richard D. Weisel ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Polymerase Chain Reaction ◽  
Growth Factor ◽  
Cell Survival ◽  
Left Ventricular ◽  
Border Zone ◽  
Lv Function ◽  
Chain Reaction ◽  
Igf I ◽  
Polymerase Chain

Background— Angiogenesis is enhanced after transplantation of vascular endothelial growth factor (VEGF)-expressing cells into a myocardial scar. Insulin-like growth factor I (IGF-I) may induce hypertrophy and inhibit apoptosis. We evaluated the effect of cell-based IGF-I and VEGF multigene therapy on left ventricular (LV) function, cell survival, and apoptosis after bone marrow cell (BMC) transplantation. Methods and Results— Female Lewis rats underwent left anterior descending ligation 3 weeks before transplantation with male donor BMC, BMC transfected with VEGF (BMC+VEGF), IGF-I (BMC+IGF-I), VEGF and IGF-I (BMC+VEGF+IGF-I), or medium without cells (control) (n=4 per group×5 groups×4 time points). Three days and 1, 2, and 4 weeks after transplantation, VEGF and IGF-I expression was quantitated by real-time polymerase chain reaction, cell survival by polymerase chain reaction for sry2, apoptosis by TUNEL staining, LV function by echocardiography and myosin heavy chain, and light chain and troponin I by Western blot. One week after transplantation, IGF-I expression in the scar and border zone was greatest in BMC+IGF-I and BMC+VEGF+IGF-I rats ( P <0.05). VEGF expression in the scar and border zone was greatest in BMC+VEGF and BMC+VEGF+IGF-I hearts ( P <0.05). Transplanted cell survival was lowest in BMC, intermediate in BMC+VEGF and BMC+IGF-I, and greatest in BMC+VEGF+IGF-I ( P <0.05). Apoptotic indices were significantly reduced in BMC+VEGF+IGF-I, BMC+VEGF, and BMC+IGF-I ( P <0.05). Two and 4 weeks after transplantation, LV ejection fraction was lowest in control, intermediate in BMC, BMC+VEGF, and BMC+IGF-I, and greatest in BMC+VEGF+IGF-I ( P <0.05). Conclusions— Transplantation of VEGF- and IGF-I-expressing BMC reduced apoptosis, maximized transplanted cell survival, and enhanced LV function. Multimodal cell-based gene therapy may maximize the benefits of cell transplantation.

scholarly journals In vivo and in vitro regulation of erythropoietin mRNA: measurement by competitive polymerase chain reaction

Blood ◽  
1993 ◽  
Vol 81 (3) ◽  
pp. 617-623 ◽  
Author(s):  
J Fandrey ◽  
HF Bunn
Keyword(s):  
Polymerase Chain Reaction ◽  
Hepatoma Cell ◽  
Hepatoma Cell Line ◽  
Mrna Levels ◽  
Chain Reaction ◽  
Human Hepatoma Cell ◽  
Competitive Polymerase Chain Reaction ◽  
Polymerase Chain

Abstract The regulation of erythropoietin (Epo) production was investigated by competitive polymerase chain reaction, a highly sensitive and accurate means of measuring Epo mRNA levels. Co-amplification of the test sample with added mutant Epo cDNA template corrects for variability in the efficiency of amplification. Epo mRNA levels were determined in tissues of normal rats and in animals with varying degrees of anemia. Reduction of the hematocrit level from 0.40 to 0.15–0.20 resulted in a 300-fold increase in kidney Epo mRNA, which comprised 80% of the total Epo mRNA versus 20% from the liver. In contrast, very low levels detected in lung and spleen were not significantly increased by anemia. The human hepatoma cell line, Hep3B, secretes high levels of Epo in response to hypoxia. This regulation is, to a large extent, transcriptional. When Hep3B cells were incubated in the presence of decreasing O2 tension from 160 to 7 mm Hg, there was a monotonic increase in Epo mRNA to 50 to 100 times the normoxic level. Hyperoxia did not suppress basal expression. When cells were incubated at a PO2 of 7 mm Hg, induction of Epo mRNA was first noted at 30 minutes and was maximal at 5 to 6 hours. After Epo mRNA was boosted by a 4-hour hypoxic incubation, cells were then exposed to normoxia, which shut off further transcription of the Epo gene. The decay of Epo mRNA levels closely followed first order kinetics with a half-life of 2 hours, an effective measurement of message stability.

scholarly journals Development of a fluorescent quantitative real-time polymerase chain reaction assay for the detection of Goose parvovirus in vivo

2009 ◽  
Vol 6 (1) ◽  
pp. 142 ◽  
Author(s):  
Jin-Long Yang ◽  
An-Chun Cheng ◽  
Ming-Shu Wang ◽  
Kang-Cheng Pan ◽  
Min Li ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Polymerase Chain Reaction Assay ◽  
Chain Reaction ◽  
Goose Parvovirus ◽  
Polymerase Chain
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