ABSTRACT
Mechanisms that direct reprogramming of differentiated somatic cells to induced pluripotent stem cells (iPSCs), albeit incomplete in understanding, are highly conserved across all mammalian species studied. Equally, proof of principle that iPSCs can be derived from domestic cattle has been reported in several publications. In our efforts to derive and study bovine iPSCs, we encountered inadequacy of methods to generate, sustain, and characterize these cells. Our results suggest that iPSC protocols optimized for mouse and human somatic cells do not effectively translate to bovine somatic cells, which show some refractoriness to reprogramming that also affects sustenance. Moreover, methods that enhance reprogramming efficiency in mouse and human cells had no effect on improving bovine cell reprogramming. Although use of retroviral vectors coding for bovine OCT4, SOX2, KLF4, cMYC, and NANOG appeared to produce consistent iPSC-like cells from both fibroblasts and cells from the Wharton's jelly, these colonies could not be sustained. Use of bovine genes could successfully reprogram both mouse and human cells. These findings indicated either incomplete reprogramming and/or discordant/inadequate culture conditions for bovine pluripotent stem cells. Therefore, additional studies that advance core knowledge of bovine pluripotency are necessary before any anticipated iPSC-driven bovine technologies can be realized.
Subject(s)
Cattle , Cellular Reprogramming , Genetic Vectors , Induced Pluripotent Stem Cells/cytology , Transcription Factors/metabolism , Animals , Cells, Cultured , Fibroblasts/cytology , Fibroblasts/metabolism , Humans , Induced Pluripotent Stem Cells/metabolism , Kruppel-Like Factor 4 , Kruppel-Like Transcription Factors/genetics , Kruppel-Like Transcription Factors/metabolism , Mice , Nanog Homeobox Protein/genetics , Nanog Homeobox Protein/metabolism , Octamer Transcription Factor-3/genetics , Octamer Transcription Factor-3/metabolism , SOXB1 Transcription Factors/genetics , SOXB1 Transcription Factors/metabolism , Transcription Factors/geneticsABSTRACT
OBJECTIVE To evaluate use of flunixin meglumine as a treatment to postpone ovulation in mares, mare fertility after flunixin meglumine treatment during estrous cycles, and effects of flunixin meglumine on function of the corpus luteum after ovulation. ANIMALS 13 healthy mares. PROCEDURES A single-blinded, placebo-controlled, crossover study was conducted. Flunixin meglumine (1.1 mg/kg, IV, q 24 h) or lactated Ringer solution (placebo treatment) was administered for 2 days to mares with a dominant follicle (≥ 35 mm in diameter) and behavioral signs of estrus. Mares then were bred by artificial insemination. Number of days to ovulation from initial detection of a follicle ≥ 30 mm in diameter, uterine edema score, and pregnancy were determined by ultrasonography; the examiner was unaware of the treatment of each mare. Serum progesterone concentrations were evaluated 5 and 12 days after ovulation by use of radioimmunoassay. RESULTS Data were available for 45 estrus cycles of the 13 mares. Number of days to ovulation from initial detection of a follicle ≥ 30 mm was not significantly affected by administration of flunixin meglumine versus the placebo. Per-cycle pregnancy rate was not significantly different between flunixin meglumine (20/24 [83%] breedings) and the placebo (13/19 [68%] breedings). Flunixin meglumine did not significantly affect behavioral signs of estrus, uterine edema, or serum progesterone concentrations. CONCLUSIONS AND CLINICAL RELEVANCE Findings did not support the use of flunixin meglumine to postpone ovulation in mares.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Clonixin/analogs & derivatives , Horses , Ovulation/drug effects , Animals , Clonixin/pharmacology , Corpus Luteum/drug effects , Cross-Over Studies , Estrous Cycle/drug effects , Estrus/drug effects , Female , Insemination, Artificial/veterinary , Pregnancy , Progesterone , Single-Blind MethodABSTRACT
A 20-month-old, 48-day pregnant female American Bulldog was presented for intermittent bloody vaginal discharge of 4-day duration. The bitch was clinically healthy with a mild neutrophilia. Transabdominal ultrasound revealed a viable foetus in the left uterine horn and fluid in the right uterine horn. She was treated with amoxicillin/clavulanic acid and re-examined at regular intervals to monitor for deterioration of the bitch or foetus. The uterine fluid progressively decreased in volume, and the puppy remained viable. A Caesarean section was performed on the bitch's expected due date, resulting in a healthy puppy. This is only the second published report of a pregnancy and concurrent pyometra with a live puppy outcome. This case is unique in that the bitch's pyometra resolved grossly during pregnancy.
