In Vitro Buffalo Embryo Production

News source: The Philippine Council for Agriculture, Forestry and Natural Resources Research and Development
For further information, Duran, D.H., et al. Pregnancies from in vitro produced and vitrified buffalo embryos after embryo transfer. Highlights 2001, PCARRD, Los Banos, Laguna 2002, pp. 155-156., 2002-03-01

In Vitro Production

Technology for in vitro (outside the animal body) embryo production (IVEP) involves techniques of in vitro oocyte maturation and in vitro fertilization (IVM/IVF). Immature buffalo oocytes are aspirated from 2 to 8 mm surface follicles of slaughtered animals, using a 18 G needle attached to a 10 ml disposable plastic syringe containing 3% Fetal Bovine Serum in Phosphate Buffered Saline. The selected oocytes are allowed to mature in a culture medium (e.g., TCM-199).

Fertilization of Mature Oocytes

After 23 - 24 hours of in vitro maturation, mature oocytes are fertilized in vitro using frozen-thawed semen. Fertilized oocytes are further cultured in the maturation medium until the embryos reach the morula, early blastocyst or expanded blastocyst stages. At these stages, good quality embryos are ready for transfer to recipient female water buffaloes.

The IVEP makes possible the use of animal genetic resources from abroad in countries which lack highly productive breeds. For instance, salvaged superior genetic materials from India may be used through IVEP by other countries. This eliminates wastage of this important resource, reduces the risk of disease transmission, and avoids the high cost of importing live animals.

Pregnancy Rate

The viability of the technique has been proven by a confirmed pregnancy rate of 14.7% among recipient animals. The highest rate of pregnancy was 25%, achieved with embryos vitrified and transferred at the early blastocyst stage, while 18.75% was achieved at the expanded blastocyst stage, and 14.29% was achieved at the morula stage. Embryo age, and recipient post-estrus synchrony suggested a + one day safety timing for embryo transfer. Continuous refinement of the technique is still in progress to improve efficiency.

Download the PDF. of this document, 28,093 bytes (27.4 KB).