A review on reproductive biotechnologies for conservation of endangered mammalian species.

Cryopreservation of gametes and embryos of non-domestic species

Embryo technology in conservation efforts for endangered felids.

Methods for preparing cell lines that contain artificial chromosomes, methods for preparation of artificial chromosomes, methods for purification of artificial chromosomes, methods for targeted insertion of heterologous DNA into artificial chromosomes, and methods for delivery of the chromosomes to selected cells and tissues are provided. Also provided are cell lines for use in the methods, and cell lines and chromosomes produced by the methods. In particular, satellite artificial chromosomes that, except for inserted heterologous DNA, are substantially composed of heterochromatin, are provided. Methods for use of the artificial chromosomes, including for gene therapy, production of gene products and production of transgenic plants and animals are also provided.

Artifical chromosomes, uses thereof and methods for preparing artificial chromosomes

Somatic cell nuclei of giant pandas can dedifferentiate in enucleated rabbit ooplasm, and the reconstructed eggs can develop to blastocysts. In order to observe whether these interspecies cloned embryos can implant in the uterus of an animal other than the panda, we transferred approximately 2300 panda-rabbit cloned embryos into 100 synchronized rabbit recipients, and none became pregnant. In another approach, we cotransferred both panda-rabbit and cat-rabbit interspecies cloned embryos into the oviducts of 21 cat recipients. Fourteen recipients exhibited estrus within 35 days; five recipients exhibited estrus 43‐48 days after embryo transfer; and the other two recipients died of pneumonia, one of which was found to be pregnant with six early fetuses when an autopsy was performed. Microsatellite DNA analysis of these early fetuses confirmed that two were from giant panda-rabbit cloned embryos. The results demonstrated that panda-rabbit cloned embryos can implant in the uterus of a third species, the domestic cat. By using mitochondrial-specific probes of panda and rabbit, we found that mitochondria from both panda somatic cells and rabbit ooplasm coexisted in early blastocysts, but mitochondria from rabbit ooplasm decreased, and those from panda donor cells dominated in early fetuses after implantation. Our results reveal that mitochondria from donor cells may substitute those from recipient oocytes in postimplanted, interspecies cloned embryos. developmental biology, early development, embryo, implantation, pregnancy

https://bioone.org/journals/Biology-of-Reproduction/volume-67/issue-2/biolreprod67.2.637/Interspecies-Implantation-and-Mitochondria-Fate-of-Panda-Rabbit-Cloned-Embryos1/10.1095/biolreprod67.2.637.pdf

Interspecies implantation and mitochondria fate of panda-rabbit cloned embryos.

The domestic cat may be used as a model for developing assisted reproduction techniques including in vitro fertilization (IVF), embryo culture, cryopreservation and embryo transfer (ET) for application to threatened and endangered species of non-domestic cats. Interoestrous domestic cats were injected with a total of 1.0-6.0 mg follicle-stimulating hormone (FSH) daily for 4 days and with 100 iu human chorionic gonadotrophin (hCG) on day 5. Follicular oocytes recovered at 26 +/- 1 h after hCG were co-incubated for 4-6 h at 38 degrees C in 5% CO2 with spermatozoa (1-2 x 10(5) motile spermatozoa ml-1) collected by artificial vagina. To determine the timing of sperm penetration and early fertilization events in vitro, oocytes were fixed and examined at intervals from 0.5 to 10 h after sperm exposure. The penetration rate of metaphase II (MII) oocytes at 0.5-3 h was equivalent to that at 3-6 h (95 versus 96%). Second polar body extrusion, pronuclear formation and apposition were observed at 2, 6-8 and 10 h, respectively. Simple (Tyrode's) and complex (F-10, M-199 and CMRL-1066) culture media with 10% fetal calf serum were compared for their ability to support development to the morula or blastocyst stage during culture periods of 96-168 h after IVF. The average number of cells per embryo was similar (P > 0.05) in the various media at each interval except that CMRL-1066 reduced (P or = 12 embryos (11/26) than if < 12 embryos were transferred (6/23).(ABSTRACT TRUNCATED AT 400 WORDS)

In vitro fertilization in domestic and non-domestic cats including sequences of early nuclear events, development in vitro, cryopreservation and successful intra- and interspecies embryo transfer.

Cumulus-oocyte complexes of domestic cats were classified by morphology of ooplasm (A = good, B = fair, C = poor) and cultured for 24 h in TCM 199 with gonadotrophins (eCG, FSH, hCG or FSH/hCG). More of type A oocytes (52%) underwent in vitro maturation (IVM) than of type B (41%) or type C (17%). The gonadotrophin source did not affect frequency of IVM of type A (50-53%) or type B (38-44%) oocytes, but IVM of type C oocytes in hCG or FSH/hCG (27%/19%) was about double that in eCG or FSH alone (13%/10%). After IVF, frequency of cleavage for type A (54%), B (41%) and C (26%) oocytes was similar to the IVM frequency of the equivalent type. After 7 days, development to the morula (M) stage in vitro was similar among types (47-58%); however, higher percentages of type A and B oocytes developed to blastocysts (Bl), 31% and 29%, respectively, than of type C (15%). After transfer of day 5 (n = 70) or 6 (n = 32) M and Bl to day 4 or 5 recipients in trial 1 (n = 4) and 2 (n = 3), respectively, the three recipients in trial 2 gave birth to four live kittens. Development in vitro to M of IVM/IVF embryos frozen in propanediol plus sucrose during early cleavage was similar (64-69%) to that of cohort controls (64%), but Bl formation was reduced (13-17% versus 32%). Damage to the zona pellucida after plunging into liquid nitrogen at -30 degrees C was higher (11%) than that of the embryos cooled at 10 degrees C min-1 from -30 degrees C to -150 degrees C before storage (2%).

In vitro and in vivo development of embryos produced by in vitro maturation and in vitro fertilization of cat oocytes

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In vitro fertilization in domestic and non-domestic cats including sequences of early nuclear events, development in vitro, cryopreservation and successful intra- and interspecies embryo transfer.

In vitro and in vivo development of embryos produced by in vitro maturation and in vitro fertilization of cat oocytes