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Studies on developmental potentials of bisected mouse embryos in vitro and in vivo.
1985
Nakagawa A. | Takahashi Y. | Kanagawa H.
Development to blastocysts of one- to two-cell equine embryos after coculture with uterine tubal epithelial cells
1993
Ball, B.A. | Brinsko, S.P. | Thomas, P.G.A. | Miller, P.G. | Ellington, J.E.
Development of 1- to 2-cell in vivo fertilized equine embryos cultured with or without uterine tubal epithelial cells (UTEC) was studied. One- to 2-cell embryos (n = 26) were collected surgically from the uterine tubes of pony mares 1 day after ovulation. Four- to 8-cell embryos (n = 9) were collected 2 days after ovulation. Presumptive zygotes and 2-cell embryos were cultured with (n = 17) or without (n = 9) UTEC, and all 4- to 8-cell embryos were cocultured with UTEC as positive controls. Uterine tubal epithelial cells were used as cell suspensions within 2 weeks after initiation of cultures. Embryos were cultured to blastocysts or until the embryo had morphologic degeneration. Six presumptive zygotes failed to cleave in vitro. Development to blastocysts of 1-cell (4 of 11) and 2-cell (2 of 6) embryos cocultured with UTEC was similar. Coculture of 1- to 2-cell embryos with UTEC significantly (P = 0.05) improved development to blastocysts, compared with culture in medium alone (35 vs 0%, respectively); however, development to blastocysts of 1- to 2-cell embryos cocultured with UTEC was less (P < 0.025) than that of 4- to 8-cell embryos cocultured with UTEC (35 vs 89%, respectively).
显示更多 [+] 显示较少 [-]Use of polymerase chain reaction to detect porcine parvovirus associated with swine embryos
1994
Gradil, C.M. | Harding, M.J. | Lewis, K.
The role of porcine parvovirus (PPV) in inducing reproductive failure in swine has been extensively documented. However, information is not available as to the risk of ppv transmission by embryo transfer. Using the polymerase chain reaction (PCR) technique, PPV-specific DNA was detected in association with 4-day-old porcine embryos incubated in vitro in the presence of NADL-8 strain of PPV, despite attempts to rid the embryos of virus by either washing or treatment with pronase or trypsin. The presence of PPV in embryos collected from acutely infected swine was not detected by PCR, although PPV DNA was detected in the proximal portion of the reproductive tract during the early stages of infection. Viral-specific nucleic acid was not detected in embryos transferred from infected donors to seronegative recipients and retrieved and assayed on the 15th and 32nd days of gestation. Results of the use of PCR to detect PPV associated with swine female reproductive tract and embryos ascribe minimal risk to the transmission of PPV to seronegative recipients through embryo transfer.
显示更多 [+] 显示较少 [-]In vitro viability of mouse oocytes vitrified in an ethylene glycol-based solution
1998
Bautista, J.A.N. (Hokkaido Univ., Sapporo (Japan)) | Pena, E.C.D. | Katagiri, S. | Takahashi, Y. | Kanagawa, H.
Ovulated mouse oocytes denuded of their cumulus cells, were vitrified in a solution containing 7 M ethylene glycol as the sole cryoprotectant using one or two steps of exposure before vitrification and were diluted in 1 M sucrose solution in 5 or 10 min after warming. The results proved that the viability of oocytes are detrimentally affected by exposure to the vitrification solution even without vitrification. At 5 min dilution time, the two-step exposure was superior to the one-step in terms of the post-warming recovery rate of vitrified oocytes with normal morphology and their subsequent development to the blastocyst stage (p0.001) after fertilization in vitro. At 10 min dilution time, no significant difference between one or two-step exposure was found. The effect of the addition of 0.5 M sucrose to the vitrification solution was also determined and did not result in a significant improvement in the viability of oocytes vitrified in one-step and diluted for 10 min. In conclusion, the results in this study indicate that oocytes can be vitrified with 7 M ethylene glycol as the sole cryoprotectant in the vitrification solution, and that the recovery of normal oocytes after one-step exposure in the vitrification solution can be improved by 10 min dilution time. However, the improvement in the recovery rate of oocytes with normal morphology and their subsequent developmental in vitro was not improved by the addition of 0.5 M sucrose to the vitrification solution
显示更多 [+] 显示较少 [-]Differentiative potential of a mouse parthenogenetic embryonic stem cell line revealed by embryoid body formation in vitro
1998
Park, J.I. (Hokkaido Univ., Sapporo (Japan)) | Yoshida, I. | Tada, T. | Takagi, N. | Takahashi, Y. | Kanagawa, H.
The in vitro differentiative potential of mouse parthenogenetic (PG) embryonic stem (PGES) cells were investigated in the formation of embryoid bodies (EBs). EBs derived from PGES cells retarded in growth and showed restricted differentiation compared to their fertilized counterpart. In chimeric EBs from the aggregation of PGES and fertilized ES cells, morphological examination revealed that PGES cells were reduced in their population and distributed in endodermal layer as culture periods proceeded. These findings were comparable to those in aggregation chimeras of fertilized and PG embryos, and suggest that the differentiation of PGES cells in vitro is restricted in the formation of EBs
显示更多 [+] 显示较少 [-]Present status of embryo transfer in water buffalo
1989
Ocampo, M.B. (Hokkaido Univ., Sapporo (Japan). Faculty of Veterinary Medicine) | Ocampo, L.C. | Rayos, A.A. | Kanagawa, H.
Effects of cycloheximide treatment on in-vitro development of porcine parthenotes and somatic cell nuclear transfer embryos
2003
Diaz, M.A.M. (Hokkaido Univ., Sapporo (Japan)) | Suzuki, M. | Kagawa, M. | Ikeda, K. | Takahashi, Y.
Effect of activation treatments of recipient oocytes on subsequent development of bovine nuclear transfer embryos
2003
Atabay, E.C. (Hokkaido Univ., Sapporo (Japan)) | Katagiri, S. | Nagano, M. | Takahashi, Y.
Trisomy 8 does not affect differentiative potential in a murine parthenogenetic embryonic stem cell line
1998
Park, J.I. (Hokkaido Univ., Sapporo (Japan)) | Yoshida, I. | Tada, T. | Takagi, N. | Takahashi, Y. | Kanagawa, H.
Murine parthenogenetic embryonic stem (ES) cell lines expressing lac zeta reporter gene were isolated after co-transfection with lac zeta reporter gene (pENL) and neoR gene (pSTneo) to TMA-48P cell line of 129/Sv origin. Karyotype analyses showed that all of four transfected cell lines examined contained 41 chromosomes with trisomy 8. Bacterial neoR transgene required for G418 selection were integrated into several chromosomes including chromosome 8. Histological studies of teratomas formed in syngenic mice and embryoid bodies grown in vitro showed that the differentiative potential remained almost identical in chromosomally normal parental cell line and its derivative cell lines trisomic for chromosome 8
显示更多 [+] 显示较少 [-]Эффективность криоконсервации эмбрионов крупного рогатого скота с использованием в качестве криопротекторов этиленгликоля и сахарозы
2008
Golubets, L.V. | Starovojtova, M.P. | Zanevskaya, E.K., Grodno State Agrarian Univ. (Belarus)
Investigation of the efficiency of ethylene glycol and sucrose application in the capacity of cryoprotectors for cryopreservation of cattle embryos and their thawing in the conditions of application of saline solutions (as their dissolution medium) and nutritive media with a various structure was realized in the conditions of the Republic of Belarus. Research results showed that application of ethylene glycol in concentration 1,5 M and sucrose in concentration 1,0M proved to be the most effective. Regardless of the applied media the average safety of embryos was 93,8-96%, and acceptability - 59,3-62,5%. Peculiar feature of ethylene glycol use as cryoprotector for preservation of cattle embryos was that it could be quickly absorbed by a cell and quickly deduced from it. It made it possible to realize embryo transfer immediately after thawing. Application of ethylene glycol and sucrose as cryoprotectors could considerably simplify the procedure of transplantation of the frozen-thawed embryos, practically reducing it to a procedure of artificial insemination
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