PARENT SESSION
Oocyte Development
78
AMINO ACID REQUIREMENTS FOR MATURATION OF RHESUS MONKEY OOCYTES IN CULTURE.
Zheng, Ping1,3, Bavister, Barry2,3, Ji, Weizhi1,3, 1 3 2
ABSTRACT- To investigate the optimal amino acids(AAs) supplement for in vitro maturation (IVM) of rhesus monkey oocytes, a total of 329 cumulus-oocyte complexes (COCs) were collected from follicles (≥1000
m) of unstimulated females. The COCs were cultured in either mCMRL-1066, or in one of eight chemically-defined media (modified Basic Medium-5 [mBM-5] with glucose [5.5 mM], pantothenic acid [0.1 mM] and different amino acid formulations) as follows: 1) mBM-5; 2) mBM-5 + glutamine (GLN, 0.2 mM); 3) mBM-5 + 11 AAs as in HECM-6; 4) mBM-5 + Eagle's nonessential amino acids (NEA); 5) mBM-5 + NEA with GLN (0.2mM); 6) mBM-5 + Eagle's essential amino acids (EA) with 0.2mM GLN; 7) mBM-5 + EA without GLN; 8) mBM-5 + Eagle's 20 AAs with 0.2mM GLN; and 9) mCMRL-1066 (control). All IVM media contained FSH (5g/ml), LH (10g/ml), P4 (3g/ml), and E2 (100 ng/ml). All mature oocytes (MII) were subjected to the same fertilization and embryo culture procedures. The maturation rate (MII) in treatment 5 (74%) was significantly higher (P<0.05) than those in treatments 1 (42%), 2 (46%), 3 (46%), 4 (53%), 7 (34%), and 9 (34%), but not significantly higher than in treatments 6 (57%) and 8 (50%). No significant differences were found among the 9 groups in terms of fertilization and cleavage rates (P>0.05). However, significant differences arose when embryos developed beyond 8-cells: the response in treatment 5 (65%) was higher than in treatments 1 (33%), 6 (24%) and 7 (38%). Treatment 8 (67%) was also higher than treatments 1 and 6. No differences existed among the other groups (2, 53%; 3, 42%; 4, 55%; 9, 50%). For further embryo development, a greater percentage (P<0.05) of morulae/blastocysts was obtained in group 8 (67%) than in groups 1 (6.7%), 2 (29%), 4 (20%), 5 (23%), 6 (14%), 7 (0), and 9 (25%). The ratio of morulae/blastocysts was also higher in group 3 (0.35) than groups 1, 6 and 7. Only group 8 supported much higher (P<0.05) blastocyst formation (33%) than any other treatments (0, 5.8%, 5.8%, 0, 0, 0, 0, and 0 in treatments 1-7 and 9, respectively). We conclude that: 1) the inclusion of certain AAs in the IVM medium can significantly increase the nuclear maturation and embryo developmental potentials of oocytes. 2) Eagle's 20 AAs with GLN is more efficient than the other AA formulations for IVM of rhesus oocytes, and its beneficial effects may be mainly attributed to NEA with GLN.
KEY WORDS: Amino acids, Oocyte in vitro maturation, Rhesus monkey