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"Pengawetan fungsi reproduksi dengan simpan beku oosit matur tidak dapat dilakukan pada pasien kanker, sehingga simpan beku oosit imatur menjadi pilihan. Simpan beku oosit imatur masih terkendala angka maturasi dan fertilisasi yang rendah. Penelitian ini bertujuan untuk mengidentifikasi pengaruh vitrifikasi sel kumulus granulosa sebagai representasi maturitas dan kualitas oosit matur dan imatur. Penanda maturitas adalah GDF-9, BMP-15, FSHR, LHR, dan koneksin-37, sedangkan kaspase-3 dan survivin sebagai penanda kualitas. Kadar protein total dan protein spesifik penanda maturitas (FSHR, LHR) dan kualitas (kaspase-3) oosit juga diteliti untuk memperoleh gambaran komprehensif pascavitrifikasi.Disain penelitian adalah true eksperimental in-vitro, dilakukan dari bulan Juli 2020 sampai Februari 2022. Subjek penelitian adalah pasien yang menjalani program fertilisasi in-vitro di klinik bayi tabung Morula IVF, RSIA Bunda Jakarta. Sampel sel kumulus-granulosa dari 38 pasien dengan diagnosis sindrom ovarium polikistik (SOPK) dianalisis. Prosedur vitrifikasi dilakukan dengan memajan sel kumulus-granulosa pada medium ekuilibrasi (VS1, 15% etilen glikol) selama 5 menit dan medium vitrifikasi (VS2, 15% etilen glikol, 15% dimethyl sulfoxide dan 0,5 M sukrosa) selama 30 detik. Sel kumulus-granulosa dimasukkan secara cepat ke nitrogen cair (suhu -196 oC). Setelah 5 menit, sampel dihangatkan dengan memajankan larutan sukrosa bertingkat 0,5 M, 0,25 M, 0.125 M. Metode real-time quantitative polimerase chain reaction (rt-qPCR) digunakan untuk mengukur ekspresi relatif dan sandwich Enzyme-linked Immunosorbent Assay (ELISA) untuk mengukur kadar protein. Analisis data menggunakan T-independent atau Mann-Whitney. Analisis ekspresi relatif gen target level mRNA menggunakan rumus Pfaffl.Didapatkan kadar protein total yang tidak berbeda bermakna antara sel kumulus-granulosa oosit matur dan imatur (30 µg/mL dan 12,5 µg/mL, nilai p > 0,05). Kadar protein spesifik FSHR, LHR dan Kaspase-3 juga tidak berbeda bermakna antara kelompok oosit matur dan imatur (masing-masing adalah 0,47 µg/mL dan 0,48 µg/mL, 0,1 µg/mL dan 0,09 µg/mL, 0.51 µg/mL dan 0.58 µg/mL, nilai p > 0,05). Tidak terdapat perbedaan bermakna ekspresi relatif BMP-15, LHR dan koneksin-37 pascavitrifikasi pada kelompok sel kumulus-granulosa oosit matur dan imatur (masing-masing 1,6 dan 1,4 kali untuk BMP-15, 2,3 dan 2,3 kali untuk LHR, 0,9 dan 0,9 kali untuk koneksin-37, nilai p < 0,01). Didapatkan penurunan bermakna ekspresi relatif GDF-9 dan FSHR pada kelompok sel kumulus-granulosa oosit matur dan imatur (masing-masing 0,2 dan 0,1 kali untuk GDF-9, 0,3 dan 0,02 kali untuk FSHR, nilai p < 0,01). Ekspresi relatif gen penanda kualitas oosit matur dan imatur yaitu survivin dan kasapase-3 tidak berubah pascavitrifikasi (1,3 dan 1,2 kali untuk survivin, 0,7 dan 0,8 kali untuk kaspase-3, nilai p > 0,05). Analisis kadar protein FSHR, LHR, dan kaspase-3 pascavitrifikasi menunjukkan bahwa vitrifikasi tidak mengubah ekspresi gen sel kumulus granulosa oosit matur dan imatur (masing-masing adalah 0,4;0,4 µg/mL dan 0;4;0,5 µg/mL untuk FSHR, 0,1;0,1 µg/mL dan 0,1;0,1 µg/mL untuk LHR, 1,9;1,5 µg/mL dan 1,7;1,6 µg/mL untuk kaspase-3, nilai p > 0,05).Disimpulkan simpan beku sel kumulus-granulosa dengan metode vitrifikasi tidak mengubah ekspresi gen penanda maturitas dan kualitas oosit kecuali GDF-9 dan FSHR. Vitrifikasi terbukti aman untuk mempertahankan viabilitas sel kumulus-granulosa sebagai representasi kesintasan oosit matur dan imatur.

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Fertility preservation through mature oocytes cannot be recommended for cancer patients and immature oocytes is preferable. However, immature oocyte vitrification remains unfavorable due to low number of mature oocytes as well as low fertilizability post-warming. This study aimed to investigate the effect of cumulus-granulosa cells' vitrification on maturity and quality-associated markers of mature and immature oocytes as an indirect assessment. Expression of GDF-9, BMP-15, FSHR, LHR, and Connexin-37 genes which represent oocyte maturity and oocyte quality (caspase-3 and survivin) were analyzed post-warming. Protein total and specific proteins including FSHR, LHR, and Caspase-3 were also measured to comprehend the effect of vitrification.

This was an in-vitro experimental study conducted from Juli 2020–February 2022 in Morula IVF Jakarta Clinic. A total of 38 cumulus-granulosa cell samples from infertile women diagnosed with polycystic ovary syndrome (PCOS) were analyzed. Vitrification was commenced by exposing the cells sample to the equilibration medium containing 15% EG for 5 minutes followed by exposing the cells to vitrification medium (15% EG and 15% DMSO) for 30 seconds. Cell samples were subsequently immersed in liquid nitrogen (-196 0C) for 5 minutes. Warming procedure using sucrose solution (0.5 M, 0.25 M, 0.125 M) was performed consecutively. Real-time quantitative polymerase chain reaction (rt-qPCR) dan sandwich ELISA was utilized to measure relative expression and concentration of both total and specific protein. Data analysis was performed using T-independent or Mann-Whitney and Pfaffl formulation for fold-changes measurement.

This study found that concentration of both total (30 µg/mL vs. 12.5 µg/mL, p > 0.05) and specific proteins was similar between cumulus-granulosa cells of mature and immature oocytes (FSHR: 0.47 µg/mL vs. 0.48 µg/mL , LHR: 0.1 µg/mL vs. 0.09 µg/mL, and caspase-3: 0.51 µg/mL and 0.58 µg/mL, respectively, p > 0.05). Relative expression of maturity-associated genes such as BMP-15, LHR and connexin-37 post-warming did not significantly different in either cumulus-granulosa cells obtained from mature or immature oocytes (BMP-15: 1.6 vs. 1.4 fold, LHR: 2.3 vs. 2.3 fold, connexin-37: 0.9 vs. 0.9 fold, p > 0.05). However, we observed decreased relative expression of GDF-9 and FSHR post-warming in either cumulus-granulosa cells obtained from mature or immature oocytes (GDF-9: 0.2 vs 0.1 fold, and FSHR: 0.3 vs. 0.02 fold, p < 0.01, respectively). In protein level, concentration of FSHR, LHR, and caspase-3 as well as protein total post-warming did not significantly different than that of fresh condition (FSHR: 0.4 vs. 0.4 µg/mL and 0.4 vs. 0.5 µg/mL, LHR: 0.1 vs. 0.1 µg/mL dan 0.1 vs. 0.1 µg/mL, and caspase-3: 1.9 vs. 1.5 µg/mL and 1.7 vs. 1.6 µg/mL, repectively, p > 0,05).

This study concluded that vitrification is a safe procedure for fertility preservation which is able to preserve cumulus-granulosa cells viability as a representative of mature and immature oocytes survival post-warming."

"The book is the third in a series of timely and indispensable books on fertility preservation for cancer patients—the first one focused on advances in basic science research and the second one offered ethical, legal, and social perspectives on the theme. This book elucidates the latest practices and emerging treatments in oncofertility and provides necessary information on the successes, risks, and limitations of fertility preserving technologies. Authoritative and insightful, written by an impressive multi-disciplinary cadre of specialists, this book is a valuable up-to-date resource for all those practicing in this demanding field."

"Tujuan: Untuk mengetahui efektifitas, kenyamanan, dan toksisitas dari alat testicular shield buatan sendiri pada preservasi fertilitas pasien di Instalasi Radioterapi pada Rumah Sakit pusat rujukan nasional di Indonesia.Metode: Penelitian deskriptif analitik, dilakukan perhitungan dosis pada TPS dan Film Dosimetry, Pemeriksaan laboratorium hormon FSH, LH, dan Testosteron pre dan post radiasi serta penilaian toksisitas kulit daerah skrotum paska radiasi dan kenyamanan penggunaan pada pasien kanker abdominopelvis yang menjalani terapi radiasi.Hasil: Total terdapat 6 pasien yang menyelesaikan proses perencanaan radiasi dengan 5 pasien berhasil menyelesaikan tatalaksana radiasi. Didapatkan Dosis rerata testis 0,8 Gy dan rerata dosis testis dibandingkan dosis preskripsi sebesar 1,8% dengan pengurangan dosis sekitar 80%. Terdapat peningkatan hormon FSH (63,5%), LH (32,2%), dan penurunan Testosteron (3,7%). Panggunaan testicular shield dapat mencegah terjadinya hipogonadisme primer akibat radiasi. Rerata skor kenyamanan 9,4 dengan tanpa ditemukan peningkatan toksisitas kulit paska radiasi.Kesimpulan: Alat testicular shield yang dibuat pada studi ini terbukti memiliki efektitas yang baik untuk mengurangi dosis yang diterima testis dengan tidak menimbulkan rasa tidak nyaman saat digunakan dan tanpa penginkatan toksisitas kulit skrotum.Kata Kunci: Pasien kanker abdominopelvis, Preservasi Fertilitas Pria. Terapi Radiasi, Testicular Shield, FSH, LH, Testosteron, Kenyamanan, Toksisitas kulit.

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Purpose: To determine the effectiveness, comfort and toxicity of In-house testicular shield devices in preserving patient fertility in radiotherapy installations at national referral hospital centers in Indonesia.

Method: Descriptive analytical research, dose calculations were carried out on TPS and Film Dosimetry, laboratory examination of FSH, LH and Testosterone hormones pre and post radiation as well as assessment of skin toxicity in the skin scrotal area post radiation and comfort of use in abdominopelvic cancer patients undergoing radiation therapy.

Results: A total of 6 patients completed the radiation planning process with 5 patients successfully completing radiation treatment. The average testicular dose was 0.8 Gy and the average testicular dose compared to the prescribed dose was 1.8% with dose reduction to testis approximately 80%, After Radiotheraphy there was an increase in the hormones FSH (63,5%), LH (32,2%), and a decrease in Testosterone (3.7%). The use of a testicular shield can prevent primary hypogonadism due to radiation. The mean comfort score was 9.4 without escalation in skin toxicity.

Conclusion: The testicular shield device created in this study was scientifically proven to have good effectiveness reducing dose received by testis with excellent comfort and without escalation in scrotal skin toxicity."