Skripsi

Pengaruh Pemberian Kalsium untuk Meningkatkan Maturasi Oosit Mencit (Mus musculus) dalam Persiapan In Vitro Fertilization (IVF) = Effect of Calcium Administration on Increasing Oocyte Maturation of Mice (Mus musculus) in Preparation for In Vitro Fertilization (IVF).

Latar belakang: Infertilitas adalah masalah kesehatan besar yang memengaruhi pasangan usia subur sebanyak 10 - 16% di dunia dan 10 - 15% di Indonesia pada tahun 2010. Salah satu tatalaksananya adalah teknologi reproduksi berbantu (TRB), yaitu in vitro fertilization (IVF). Namun, tingkat keberhasilannya rendah (47,5%) dan biayanya mahal. Selain disebabkan oleh sperma, kegagalan fertilisasi juga disebabkan oleh tingginya proporsi oosit yang imatur. Oosit imatur dapat dimaturasi secara in vitro (MIV), tetapi tingkat keberhasilannya juga cukup rendah (36%) karena beberapa kondisi yang dapat menyebabkan oosit imatur sulit dimaturasi. Beberapa penelitian telah membuktikan bahwa kalsium berperan dalam fisiologi oosit, mulai dari maturasi, aktivasi, hingga fertilisasinya. Penggunaan kalsium dalam meningkatkan keberhasilan aktivasi oosit juga telah diterapkan beberapa tahun belakangan ini. Namun, belum terdapat penelitian yang mempelajari potensi pemberian kalsium dalam meningkatkan maturasi oosit. Oleh karena itu diperlukan penelitian untuk mengungkapkan potensi kalsium sebagai strategi meningkatkan keberhasilan maturasi in vitro (MIV) oosit imatur. Metode: Penelitian analitikal ini menggunakan desain eksperimental dengan menggunakan data primer dari percobaan hewan uji mencit (Mus musculus). Sampel oosit segar yang diperoleh dari mencit betina galur DDY berusia 6 – 8 minggu didenudasi, lalu dinilai maturitasnya menggunakan mikroskop stereo. Oosit tergolong matur (n = 161) jika telah mencapai tahap meiosis II yang ditandai dengan keberadaan badan polar pertama, sedangkan oosit tergolong imatur (n = 334) jika masih berada pada tahap meiosis I (tidak terdapat badan polar pertama) atau terdapat vesikel germinal (VG). Selanjutnya, oosit imatur dibagi ke dalam kelompok perlakuan (n = 106) yang diberikan 5 µM kalsium ionofor (CaI) selama 5 menit dan kelompok kontrol (n = 228) tanpa pemberian kalsium. Kemudian, kedua kelompok tersebut menjalani proses MIV, lalu dinilai maturitasnya. Data maturasi oosit dianalisis dengan uji Chi-square menggunakan SPSS. Hasil: Proporsi oosit yang berhasil dimaturasi lebih tinggi pada kelompok maturasi oosit imatur yang didahului pemberian kalsium (perlakuan) (45,3%) dibandingkan dengan kelompok maturasi oosit imatur tanpa didahului pemberian kalsium (kontrol) (32%), dengan perbedaan sebesar 13,3% yang signifikan (p < 0,05). Proporsi oosit matur juga lebih tinggi pada kelompok perlakuan (45,3%) dibandingkan dengan kelompok sampel oosit segar (32,5%), dengan perbedaan sebesar 12,8% yang signifikan (p < 0,05). Namun, tidak ditemukan perbedaan yang signifikan antara proporsi oosit matur pada kelompok kontrol dan sampel oosit segar (p > 0,05). Kesimpulan: Pemberian kalsium dapat meningkatkan maturasi in vitro oosit imatur dibandingkan dengan oosit imatur yang tidak diberi kalsium. Dengan demikian, pemberian kalsium berpotensi menjadi strategi baru dalam meningkatkan keberhasilan maturasi in vitro (MIV) oosit. Penelitian lebih lanjut dan uji klinis perlu dilakukan agar strategi ini dapat diterapkan pada praktek IVF.
Kata kunci: Kalsium, Maturasi Oosit, Maturasi In Vitro (MIV), In Vitro Fertilization (IVF), Mencit (Mus musculus), Oosit Imatur



Introduction: Infertility is a major health problem, affecting childbearing-age couples 10 - 16% worldwide and 10 - 15% in Indonesia in 2010. One of the treatments is assisted reproductive technology (ART), such as in vitro fertilization (IVF). However, the success rate is low (47.5%) and the cost is high. Aside from sperm factor, fertilization failure is also caused by the high proportion of immature oocytes. Immature oocytes can be matured in vitro (IVM), but the success rate is also low (36%) due to several conditions that can make it difficult for immature oocytes to mature. Several studies have proven that calcium plays a role in oocyte physiology, from maturation to activation, and fertilization. Calcium has been the most widely used for artificial oocyte activation. However, no research studies the potential of calcium supplementation in increasing oocyte maturation. Therefore, research is needed to reveal the potential of calcium as a strategy to increase the success of in vitro maturation (IVM) of immature oocytes. Method: This analytical study used an experimental design using primary data from experimental animals – mice (Mus musculus). Fresh oocytes samples obtained from DDY female mice aged 6-8 weeks were denuded, then assessed for maturity using a stereo microscope. The oocytes were classified as mature (n = 161) if they had reached the stage of meiosis II which was characterized by the presence of the first polar body, while immature oocytes are classified (n = 334) if they were still in the meiosis I stage (no first polar body) or there were germinal vesicles (GV). Next, immature oocytes were divided into a treatment group (n = 106) which was given 5 µM calcium ionophores (CaI) for 5 minutes, and a control group (n = 228) without calcium. Then, the two groups underwent the IVM process, and then their maturity was assessed. Oocyte maturation data were analyzed by Chi-square test using SPSS. Result: The proportion of successfully matured oocytes was higher in the immature oocyte maturation group that was preceded by calcium supplementation (treatment) (45.3%) compared to the immature oocyte maturation group without calcium supplementation (control) (32%), with a difference of 13, 3% which was significant (p < 0.05). The proportion of mature oocytes was also higher in the treatment group (45.3%) compared to the fresh oocyte sample group (32.5%), with a significant difference of 12.8% (p < 0.05). However, there was no significant difference between the proportion of mature oocytes in the control group and the fresh oocyte sample (p > 0.05). Conclusion: Calcium supplementation can increase in vitro maturation of immature oocytes compared to immature oocytes that are not given calcium. Thus, calcium supplementation has the potential to be a new strategy for increasing the success of oocyte in vitro maturation (MIV). Further research and clinical trials need to be carried out to apply this strategy to IVF practice in the clinical setting.
Keywords: Calcium, Oocyte Maturation, In Vitro Maturation (IVM), In Vitro Fertilization (IVF), Mice (Mus musculus), Immature Oocyte