Text

Interaksi Komponen Pro dan Antiapoptosis Sel Granulosa Penanggap Baik Pasca Stimulasi Ovarium Terkendali yang Dipajankan pada Medium Terkondisikan Sel Punca Mesenkim Talipusat = Interaction Between Pro- and Antiapoptotic Component Of Normo-Responders’ Granulosa Cells After Controlled Ovarian Stimulation Exposed To Umbilical Cord Mesenchymal Stem Cells Derived Conditioned Medium.

Medium terkondisikan sel punca mesenkim dari talipusat (MT SPM-TP) menghasilkan sekretom bersifat antiapoptosis yang berpotensi mencegah apoptosis sel granulosa ketika dilakukan stimulasi ovarium terkendali (SOT). Kajian ini merupakan penelitian eksperimental dengan rancangan sebelum dan sesudah perlakuan tanpa kontrol yang menggunakan biakan sel granulosa yang dipajankan pada MT SPM-TP untuk memperoleh data tampilan Bax, Survivin, nisbah Bax/survivin, GDF9 dan kadar IGF-1 di MT SPM-TP. Penelitian dilakukan di RSCM dan FKUI dari Februari 2017 hingga Desember 2018, menggunakan 23 subjek FIV penanggap baik dengan rerata umur 32,57  5,80 tahun. Persentase oosit matang per total jumlah oosit adalah 79,57%. Ditemukan perbedaan bermakna secara statistik pada tampilan Bax pasca dipajankan pada MT SPM-TP (4,09E-7 vs. 3,74E-7, p = 0,024). Tidak ditemukan perbedaan bermakna secara statistik pada survivin (3,04E-8 vs. 2,31E-8, p = 0,571), nisbah Bax / survivin (6,98E+1 vs. 8,5E+0, p = 0.199), dan GDF9 (4,88E-5 vs. 5,98E-5, p = 0,317) pasca dipajankan pada MT SPM-TP. Terdapat peningkatan yang bermakna secara statistik pada kadar IGF-1 di dalam MT SPM-TP pasca pemajanan (2,28 vs. 3,07  1,72, p < 0,001). Uji korelasi memperlihatkan hubungan positif kuat dan bermakna secara statistik antara survivin dan GDF9 (r = 0,096, p < 0,001). Analisis kuantitatif subjek yang persentase oosit matangnya < 70% memperlihatkan penurunan tampilan Bax dan nisbah Bax/survivin serta peningkatan tampilan GDF 9 lebih nyata pada subjek yang persentase oosit matangnya sangat rendah (16.67%). Subjek dengan persentase oosit matangnya < 70% tidak memperlihatkan peningkatan tampilan survivin pasca pemajanan. MT SPM-TP terbukti dapat menekan tampilan Bax, namun tidak mampu memicu peningkatan tampilan survivin dan produksi GDF9 di sel granulosa. Meski demikian terdapat korelasi yang positif dan kuat antara tampilan survivin dengan produksi GDF9 di sel granulosa. Di bawah pengaruh MT SPM-TP, sel granulosa terbukti dapat menghasilkan IGF-1, yang berpotensi bekerja secara autokrin di sel granulosa. Hasil analisis kuantitatif individual pada subjek dengan persentase kematangan oosit yang rendah memperlihatkan adanya tanggapan sel granulosa yang beragam terhadap pajanan MT SPM-TP. Namun sel granulosa yang berasal dari subjek dengan persentase kematangan oosit terendah menunjukkan tanggapan yang lebih baik pasca pajanan pada MT SPM-TP dari aspek penurunan Bax dan peningkatan produksi GDF9.
Kata kunci: Bax, GDF 9, IGF-1, Medium terkondisikan, Survivin


Conditioned medium from umbilical cord derived mesenchymal stem cell (CM UC-MSC) produces secretomes with antiapoptotic property, which is potential to prevent granulosa cells apoptosis during controlled ovarian hyperstimulation (COH). This study was an experimental study with pre- and post-treatment design without control, using granulosa cell culture exposed to CM UC-MSCs to evaluate Bax, survivin and Bax/survivin ratio, GDF9 in granulosa cell and level of IGF-1 in CM. This study was done at RSCM and FKUI from February 2017 to December 2018, enrolling 23 IVF patients, who were categorized as normo-responder. Subject’s average age was 32.57  5.80 years old. The percentage of matange oocyte based on total oocytes collected was 79.57%. There was a significant difference on Bax expression after CM UC-MSCs exposure (4.09E-7 vs. 3.74E-7, p = 0.024). However, there was no significant changes on survivin (3.04E-8 vs. 2.31E-8, p = 0.571), ratio Bax/survivin (6.98E+1 vs. 8.5E+0, p = 0.199), and GDF9 (4.88E-5 vs. 5.98E-5, p = 0.317) after CM UC-MSCs exposure. IGF-1 level of CM was significantly rise after being exposed to CM UC-MSCs (2.28 vs. 3.07  1.72, p < 0.001). Correlation study showed a significant positive and strong correlation between changes on survivin and GDF9 (r = 0.096, p < 0.001). Quantitative analysis on subjects with mature oocyte < 70%, showed a decrease in expression of Bax, and ratio of Bax/ survivin; while the elevation of GDF9 was more obviously seen in subject with lowest oocyte maturation (16.67%). No subject with mature oocyte < 70% demonstrated elevation in survivin after CM UC-MSCs exposure. CM UC-MSC has capability to reduce antiapoptotic activity by reducing Bax expression. Unfortunately, CM UC-MSC was failed to increase survivin expression and GDF9 production in granulosa cells. However, positive and strong correlation has been demonstrated between survivin and GDF9 in granulosa cells. Moreover, granulosa cells also showed its capability to produce IGF-1 after cultured with CM UC-MSC. Individual quantitative analysis on subject with low percentage in mature oocyte showed various response to CM UC-MSC exposure. However, granulosa cells coming from subject with the lowest percentage in mature oocyte gaining more benefits after exposed with CM UC-MSC in terms of Bax reduction, and more GDF9 production.
Keywords: Bax, Conditioned medium, GDF 9, IGF-1, Survivin