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Abstrak :
ABSTRAK
Penelitian dilakukan untuk mengevaluasi penambahan antioksidan alfa tokoferol dalam media pembekuan lambat terhadap kualitas oosit domba garut hingga kriopreservasi menggunakan metode pembekuan lambat. Sebanyak 226 oosit kualitas A dan B dimatangkan sebelumnya pada media pematangan TCM-199 dengan penambahan alfa tokoferol 150 μM, kemudian diawetkan dalam media pembekuan lambat dengan penambahan alfa tokoferol 0 μM (KK), 100 μM (KP1), 150. μM (KP2), dan 200 μM (KP3). Dalam media pembekuan lambat, etilen glikol 10% dan sukrosa 0,1 M juga ditambahkan. Evaluasi oosit dilakukan setelah 7 hari disimpan dalam nitrogen cair (-196 oC), meliputi morfologi oosit dan viabilitas oosit menggunakan pewarna Hoechst dan Propidium Iodide (PI). . Berdasarkan hasil yang diperoleh, persentase oosit normal pasca kriopreservasi adalah 0 μM (67,86%), 100 μM (68,42%), 150 μM (74,58%), dan 200 μM (61,11%). Persentase oosit hidup setelah kriopreservasi adalah 0 μM (76,79%), 100 μM (80,70%), 150 μM (86,44%), dan 200 μM (70,37%). Hasil penelitian menunjukkan bahwa penambahan antioksidan alfa tokoferol tidak berpengaruh terhadap kualitas oosit pasca kriopreservasi, namun terdapat kecenderungan peningkatan kualitas seiring dengan peningkatan dosis alfa tokoferol. Pada penelitian ini penambahan 150 μM alfa tokoferol dalam media pembekuan lambat merupakan dosis terbaik dalam menjaga kualitas oosit hingga pasca kriopreservasi, walaupun tidak signifikan.
ABSTRACT
The study was conducted to evaluate the addition of alpha tocopherol antioxidants in slow freezing media on the oocyte quality of arrowroot sheep until cryopreservation using the slow freezing method. A total of 226 oocytes of A and B quality were pre-ripened on the TCM-199 ripening medium with the addition of 150 μM alpha tocopherol, then preserved in slow freezing media with the addition of alpha tocopherol 0 μM (KK), 100 μM (KP1), 150. μM (KP2) , and 200 μM (KP3). In slow freezing medium, 10% ethylene glycol and 0.1 M sucrose were also added. Oocyte evaluation was carried out after 7 days of storage in liquid nitrogen (-196 oC), including oocyte morphology and oocyte viability using Hoechst and Propidium Iodide (PI) stains. . Based on the results obtained, the percentage of normal oocytes after cryopreservation were 0 μM (67.86%), 100 μM (68.42%), 150 μM (74.58%), and 200 μM (61.11%). The percentage of live oocytes after cryopreservation was 0 μM (76.79%), 100 μM (80.70%), 150 μM (86.44%), and 200 μM (70.37%). The results showed that the addition of alpha tocopherol antioxidants did not affect the quality of post-cryopreservation oocytes, but there was a tendency to increase in quality along with the increase in alpha tocopherol doses. In this study, the addition of 150 μM alpha tocopherol in slow freezing medium was the best dose in maintaining oocyte quality until post cryopreservation, although it was not significant.

Abstrak :
Telah dilakukan penelitian tentang penambahan antioksidan glutathione ke dalam medium pembekuan lambat terhadap kualitas oosit domba garut (Ovis aries) pascakriopreservasi. Tujuan dari penelitian ini adalah untuk mengevaluasi pengaruh penambahan antioksidan glutathione dalam media pembekuan lambat dengan konsentrasi 0,5 mM; 1 mM; 1,5 mM melawan kualitas oosit domba garut. Penelitian dilakukan di Lab. Reproduksi, Pemuliaan dan Kultur Sel Hewan LIPI, Cibinong. Kriopreservasi oosit dari 136 oosit dilakukan menggunakan krioprotektan 10% etilen glikol dan sukrosa 0,1 M. Evaluasi oosit dilakukan setelah 7 hari penyimpanan dalam nitrogen cair (-196°C), meliputi: morfologi oosit dan viabilitas oosit menggunakan pewarna Hoechst dan propidium pewarna iodida. Berdasarkan hasil penelitian didapatkan persentase oosit normal normal pascakriopreservasi yaitu 0 mM (68,40%), 0,5 mM (66,98%), 1 mM (73,05%), dan 1,5 mM (80,58%). Persentase oosit yang layak pasca-kriopreservasi adalah 0 mM (68,40%), 0,5 mM (69,76%), 1 mM (75,55%), dan 1,5 mM (83,08%). Berdasarkan uji statistik ANOVA, didapatkan hasil bahwa tidak ada perbedaan yang signifikan antar kelompok perlakuan (P > 0,05), namun grafik persentase menunjukkan pola yang cenderung meningkat dengan penambahan konsentrasi antioksidan glutathione. Kesimpulan penelitian berdasarkan hasil yang diperoleh yaitu penambahan antioksidan glutathione dalam media pembekuan lambat tidak berpengaruh terhadap kualitas oosit domba garut pasca-kriopreservasi.
Research has been carried out on the addition of glutathione as an antioxidant in slow freezing medium on the quality of post-cryopreserved arrowroot sheep (Ovis aries) oocytes. The aim of this study was to evaluate the effect of adding the antioxidant glutathione in slow freezing medium with a concentration of 0.5 mM; 1 mM; 1.5 mM against arrowroot sheep oocyte quality. The research was conducted in the Lab. Animal Cell Reproduction, Breeding and Culture LIPI, Cibinong. Oocyte cryopreservation of 136 oocytes was performed using cryoprotectant 10% ethylene glycol and 0.1 M sucrose. Evaluation of oocytes was carried out after 7 days of storage in liquid nitrogen (-196°C), including: oocyte morphology and oocyte viability using Hoechst stain and propidium iodide dye. Based on the results, the percentage of post-cryopreserved normal oocytes was 0 mM (68.40%), 0.5 mM (66.98%), 1 mM (73.05%), and 1.5 mM (80.58%). . The percentage of viable post-cryopreservation oocytes were 0 mM (68.40%), 0.5 mM (69.76%), 1 mM (75.55%), and 1.5 mM (83.08%). Based on the ANOVA statistical test, it was found that there was no significant difference between the treatment groups (P > 0.05), but the percentage graph shows a pattern that tends to increased with the addition of the antioxidant glutathione concentration. The conclusion of the study based on the results obtained was that the addition of the antioxidant glutathione in the slow freezing medium had no effect on the post-cryopreservation quality of arrowroot sheep oocytes.

Abstrak :
Penelitian mengenai pengaruh penambahan antioksidan glutathione GSH pada medium maturasi oosit domba garut pascakriopreservasi dengan metode slow freezing telah dilakukan sejak Maret hingga Mei 2018. Penelitian bertujuan untuk mengetahui pengaruh penambahan GSH pada medium maturasi oosit terhadap jumlah oosit matur. Oosit dimatangkan dalam medium TCM-199 yang diberi penambahan GSH sebanyak 0 mM, 0,5 mM, 1 mM, dan 1,5 mM, kemudian dibekukan menggunakan metode slow freezing dengan suhu seeding -7oC. Oosit matur ditunjukkan oleh terbentuknya badan polar I dan atau terjadinya ekspansi sel kumulus. Data viabilitas oosit pascakriopreservasi diperoleh dengan menggunakan pewarna fluorescence Hoechst/PI, sedangkan data kualitas oosit pascakriopreservasi diperoleh berdasarkan morfologi oosit. Hasil penelitian menunjukkan penambahan antioksidan GSH pada medium maturasi oosit secara statistik berpengaruh nyata P le;0,05 pada data oosit matur. Data hasil viabilitas dan kualitas oosit pascakriopreservasi oosit hasil maturasi in vitro dengan metode slow freezing tidak berpengaruh nyata secara statistik P>0,05 antarkelompok perlakuan. Kesimpulan penelitian penambahan GSH pada medium maturasi oosit domba garut secara statistik berpengaruh positif terhadap jumlah oosit yang matur. ...... The study on the effect of the addition of antioxidant glutathione GSH on Garut sheep rsquo s oocyte maturation medium after cryopreservation with slow freezing method has been done since March to May 2018. The study was conducted to determine the effect of the addition of GSH on medium maturation towards the percentage of mature oocyte. Oocytes were matured in a TCM 199 which added GSH of 0 mM, 0,5 mM, 1 mM, and 1,5 mM, then frozen using the slow freezing method and the seeding temperature used was 7oC. The mature oocyte indicated by the formation of polar body I and or the expansion of cumulus. Data of viability oocytes after cryopreservation was obtained by using Hoechst PI fluorescence dye, whereas data of quality oocytes after cryopreservation was obtained based on oocyte morphology. The results showed that the addition of GSH antioxidant in oocyte maturation medium significantly affect P le 0,05 The result data of viability and quality of oocytes after cryopreservation of in vitro oocyte maturation by slow freezing method did not significantly affect P 0.05 statistically among treatment groups. The conclusion of the study is the addition of GSH on maturation oocyte garut sheep medium does have a positive effect statistically on the number of mature oocytes.

Abstrak :
Penelitian mengenai potensi antioksidan alfa-tokoferol terhadap tingkat maturasi dan kualitas oosit domba garut (Ovis aries) pascakriopreservasi menggunakan metode slow freezing telah dilakukan. Sebanyak 138 oosit yang memiliki kualitas A dan B dimaturasi dalam medium TCM-199 dengan penambahan antioksidan alfa-tokoferol pada konsentrasi 0 mM, 100 mM, 150 mM, dan 200 mM. Oosit hasil maturasi in vitro tersebut, kemudian dikriopreservasi serta diamati tingkat maturasi dan kualitasnya. Pengamatan tingkat maturasi dilihat berdasarkan pembentukan badan polar I. Pengamatan kualitas oosit meliputi kondisi morfologi dan viabilitas menggunakan pewarna Hoechst dan propidium iodide. Hasil penelitian menunjukkan bahwa tidak ada perbedaan signifikan antarkonsentrasi secara statistik (P > 0,05), namun terdapat kecenderungan konsentrasi 150 mM memiliki persentase tertinggi terhadap tingkat maturasi oosit in vitro (88,57%); viabilitas (82,86%); dan kondisi morfologi (82,86%). Dengan demikian, 150 mM alfa-tokoferol merupakan konsentrasi terbaik yang mampu mempertahankan stabilitas oosit pascakriopreservasi.

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The study of the potential alpha-tocopherol antioxidants towards garut sheep (Ovis aries) oocyte maturation and quality post-cryopreservation using slow freezing method has been conducted. A total of 138 oocytes which have A and B qualities, were maturating in TCM-199 medium with the addition of alpha-tocopherol antioxidants at concentrations of 0 mM, 100 mM, 150 mM, and 200 mM. Oocytes from in vitro maturation are then cryopreserved and the maturity and quality was observed. Observation of oocyte maturation was seen based on the formation of the polar body I. Observation of oocytes quality includes morphological condition and viability by Hoechst and propidium iodide dyes. This experiment showed that there was no statistically siginificant difference between concentrations (P > 0.05), but there is a tendency of 150 mM have the highest percentage of oocyte in vitro maturation (88.57%); viability (82.86%); and morphological condition (82.86%). In conclusion, presence of 150 mM alpha-tocopherol was the optimal concentration that is able to maintain the oocyte membrane stability post-cryopreservation.