Effect of Various Culture Media on Mouse 2-cell Development

Chainarong Tocharus; Jiraporn Tocharus

Authors

Chainarong Tocharus Department of Anatomy, Faculty of Medical Science, Naresuan University, Phitsanulok 65000, Thailand.
Jiraporn Tocharus Department of Anatomy, Faculty of Medical Science, Naresuan University, Phitsanulok 65000, Thailand.

Keywords:

mouse embryo, embryo development, hatched-blastocyst, co-culture

Abstract

Developing a culture system for embryos has important biotechnological implications due to the potential to produce large number of preimplantation embryos. As an initial step toward accomplishing this long-term goal, a study was conducted to determine the effects of culture media on embryo development in vitro. These studies were designed to examine the development of mouse embryos from the 2-cell to the hatched-blastocyst stages in the presence of various co-culture systems comparing with culture medium alone. In vivo fertilized 2-cell embryos were surgically collected from superovulated mice 48 hrs after mating. Embryo were randomly divided into 7 culture groups as follows: culture with TCM199+BSA, commercial 1 or commercial 2; co-culture with bovine, porcine, swamp buffalo oviduct epithelial cells or mouse granulosa cells. After 72 hrs in culture, the proportion of 2-cell embryos developed to blastocyst in commercial 2 or swamp buffalo oviductal cells were significantly higher than other culture media or co-culture. The embryos developed to hatched blastocyst were significantly higher in commercial 2 medium than swamp buffalo. Physicochemical analysis indicated that the substances in all culture media were predominantly detected at 66 kDa. These results revealed that embryos cultured in medium alone were more effective in supporting mouse embryo development than the embryos cultured in co-culture system. Therefore, the available commercial culture medium has provided a new in vitro model for embryo production research in numerous species of animals.

References

Barnes, F. L., Crombie, A., Gardner, D. K., Kausche, A., Lacham-Kaplan, O., Suikkari, A. M., et al. (1995). Blastocyst development and birth after in vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching. Human Reproduction, 10, 3243-3247.

Behr, B., Pool, T. B., Milki, A. A., Moore, D., Gebhardt, J., & Dasig, D. (1999). Preliminary clinical experience with human blastocyst development in vitro without co-culture. Human Reproduction, 14, 454-457.

Chatot, C. L., Ziomek, C. A., Bavister, B. D., Lewis, J. L., & Torres, L. (1985). An improved culture medium supports development of random-bred 1-cell mouse embryos in vitro. Journal of Reproduction and Fertility, 86, 679-688.

Davis, D. L., & Day, B. N. (1987). Cleavage and blastocyst formation by pig eggs in vitro. Journal of Animal Science, 46, 1043-1053.

Ellington, J. E., Carney, E. W., Farrel, P. B., Simkin, M. E., & Foote, R. H. (1990). Bovine 1-2-cell embryo development using a simple medium in three oviduct epithelial cell co-culture systems. Biology of Reproduction, 43, 97-104.

Freeman, M., Whitworth, M., & Hill, G. (1995). Granulosa cell co-culture enhances human embryo development and pregnancy rate following in vitro fertilization. Human Reproduction,10, 408-414.

Gardner, D. K., Lane, M., Calderon, I., & Leeton, J. (1996). Environment of the preimplantation human embryo in vivo: Metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells. Fertility and Sterility, 65, 349-353.

Gott, A. L., Hardy, K., Winston, R. M., & Leese, H. J. (1990). Non-invasive measurement of pyruvate and glucose uptake and lactate production by single human preimplantation embryos. Human Reproduction, 5, 104-108.

Hardy, K., Hooper, M. A., Handyside, A. H., Rutherford, A. J., Winston, R. M., & Leese, H. J. (1989). Non-invasive measurement of glucose and pyruvate uptake by individual human oocytes and preimplantation embryos. Human Reproduction,4, 188-191.

Huisman, G. J., Alberda, A., Laerentveld, R. A., & Zeilmaker, G. M. (1994). Implantation rates afterin vitro fertilization and transfer of a maximum of two embryos that have undergoes three to five drops of culture. Fertility and Sterility, 73, 117-122.

Jones, G. M., Trounson, A. O., Gardner, D. K., Kausche, A., Loatgis, N., & Wood, C. (1998). Evolution of a culture protocol for successful blastocyst development and pregnancy. Human Reproduction, 13, 169-177.

Krisher, R. L., Lane, M., & Bavister, B. D. (1999). Developmental competence and metabolism of bovine embryos cultured in semi-defined culture media. Biology of Reproduction, 60, 1345-1352.

Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 15, 680-685.

Langendonckt, A. V., Demylle, D., Wyns, C., Nisolle, M., & Donnez, J. (2001). Comparison of G1.2/B2.2 and Sydney IVF cleavage/blastocyst sequential media for the culture of human embryos: A prospective, randomized, comparative study. Fertility and Sterility, 76, 1023-1031.

Leese, H. J., Hooper, M. A., Edwards, R. G., & Ashwood-Smith, M. J. (1986). Uptake of pyruvate by early human embryos determined by a non-invasive technique. Human Reproduction, 1, 181-182.

Macklon, N. S., Pieters, M. H. E. C., Hassan, M. A., Jeucken, P. H. M., Eijkemans, M. J. C., & Fauser, B. C. J. M. (2002). A prospective randomized comparison of sequential versus monoculture systems for in-vitro human blastocyst development. Human Reproduction, 17, 2700-2705.

Mao, J., Wu, G., Smith, M. F., McCauley, T. C., Prather, R. S., Didion, B. A., et al. (2002). Effects of culture medium, serum type, and various concentration of follicule-stimulating hormone on porcine preantral follicular development and antrum formation in vitro. Biology of Reproduction, 67, 1197-1203.

Minami, N. (1996). Early embryonic development under oviductal influence in vitro. Animal Reproduction Science, 42, 361-369.

Minami, N., Bavister, B. D., & Iritani, A. (1988). Development of hamster two-cell embryos in the isolated mouse oviduct in organ culture system. Gamete Research,19, 235-240.

Neimer, K. E., Hoffman, D. I., & Maxson, W. S. (1993). Embryonic morphology and rate of implantation of human embryos following co-culture on bovine oviductal epithelial cells. Human Reproduction, 8, 97-101.

Pavasuthipaisit, K., Lhuangmahamongkol, S., Tocharus, C., Kitiyanant, Y., & Prempree, P. (1994). Porcine oviductal cells support in vitro bovine embryo development. Theriogenology,41, 1127-1138.

Plachot, M. R., Witworth, C. M., & Hill, G. A. (1995). Granulosa cell co-culture enhances human embryo development and pregnancy rate following in vitro fertilization. Human Reproduction, 10, 408-414.

Quinn, P., & Margalit, R. (1996). Beneficial effects co-culture with cumulus cells on blastocyst formation in a prospective trial with supernumerary human embryo. Journal of Assisted Reproduction and Genetics, 13, 9-14.

Rieger, D., Grisart, B., Semple, E., Van Langendonckt, A., Betteridge, K. J., & Dessy, F. (1995). Comparison of the effects of oviductal cell co-culture and oviductal cell-conditioned medium on the development and metabolic activity of cattle embryos. Journal of Reproduction and Fertility, 105, 91-98.

Saito, H., Hirayama, T., & Kojne, K. (1994). Cumulus mass maintains embryo quality. Fertility and Sterility, 62, 555-558.

Sirard, M. A., Lamisert, R. D., Menard, D. P., & Bedoya, M. (1985). Pregnancies after in vitro fertilization of cow follicular oocytes, their incubation in rabbit oviduct and their transfer to the cow uterus. Journal of Reproduction and Fertility, 75, 551-556.

Swain, J. E., Bormann, C. L., & Krisher, R. L. (2001). Development and viability of in vitro derived porcine blastocysts cultured in NCSU23 and G1.2/G2.2 sequential mediuim. Theriogenology, 56, 459-469.

Tarin, J. J., de Santos, M. J., de Oliveira, M. N., Pellicer, A., & Bonilla-Musoles, F. (1994). Ascorbate-supplemented media in short-term cultures of human embryos. Human Reproduction, 9, 1717-1722.

Thibault, C. (1966). In vitro culture of cow egg. Annales De Biologie Animale, Biochimie, Biophysique, 6, 159-164.

Weiner, K. E., & Cohen, J. (1990). Co-culture of mammalian embryos. Amsterdam: Elsevier Science Press.

Weimer, K. E., Cohen, J., Tucker, M. J., & Godek, R. A. (1998). The application of co-culture in assisted reproduction: 10 years of experience with human embryos. Human Reproduction, 13, 226-238.

Whittingham, D. E., & Bavister, B. D. (1974). Development of hamster eggs fertilized in vitro or in vivo. Journal of Reproduction and Fertility, 38, 489-492.