MULYOTO PANGESTU
1. EPRD- Dept. Obstetrics and Gynecology, Monash University, Monash Medical Center,Victoria, Melbourne 2.Lab. Reproductive Physiology, Jenderal Soedirman University, Purwokerto

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MICROVOLUME OF 0.1µL GAMA SLEEVED CRYOLOOPS FOR BLASTOCYST VITRIFICATION OF ASSISTED REPRODUCTIVE TECHNOLOGY PATIENTS Hanoum, Ita Fauzia; Boediono, Arief; Pangestu, Mulyoto; Haryadi, Dwi; Widad, Shofwal; Dasuki, Djaswadi
JURNAL KESEHATAN REPRODUKSI Vol 2, No 1 (2015)
Publisher : IPAKESPRO

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (181.875 KB) | DOI: 10.22146/jkr.7127

Abstract

Ita Fauzia Hanoum1,2, Arief Boediono3, Mulyoto Pangestu4,5, Dwi Haryadi1,Shofwal Widad1,2, Djaswadi Dasuki1,2 ABSTRAK Latar Belakang: Prosedur embrio vitrifikasi menggunakan alat berupa grid, straw atau cryoloop. Gama Sleeved cryoloop dibuat dan dikembangkan di klinik Permata Hati. Untuk itu, dilakukan pengamatan keberhasilan prosedur vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop.Metode: Vitrifikasi dilakukan pada blastokis dengan kualitas baik yang diperoleh pada hari ke 5 setelah fertilisasi. Inform consent telah disampaikan sebelumnya kepada pasien program bayi tabung di Klinik Permata Hati. Prosedur dilakukan dengan menggunakan media handling (GMOPS Plus; Vitrolife) embrio diinkubasi selama 1 menit; (7.5% EG (v/v); 7.5% DMSO (v/v)) selama 2-3 menit, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) selama 30 detik pada suhu ruang sebelum kemudian diletakkan di dalam cryoloop, setelah itu secara cepat cryoloop yang berisi embrio dibenamkan ke dalam nitrogen cair. Sebelum dilakukan embryo transfer (ET), embrio dihangatkan dengan cara two step technique (sucrose 0.25M) selama 2 menit dan selama 3 menit (sucrose 0.125M).Hasil: Sejumlah 97 blastokis divitrifikasi dan dihangatkan (67 pasien), dimana 91 blastokis berhasil ditransfer ke rahim ibu (93.8%). Blastokis yang tidak berhasil selamat dari prosedur penghangatan adalah blastokis dengan kerusakan lebih dari 50%. Diperoleh kehamilan klinis 43.3% sedangkan angka implantasi adalah 37.4%. Sampai saat ini, dilaporkan 20 kelahiran (23 bayi) dari program vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop, sementara 5 kehamilan masih berlangsung. Satu kehamilan dilaporkan gugur pada usia kehamilan yang masih sangat awal, dua keguguran pada usia kehamilan 12 minggu dan satu bayi lahir meninggal karena kelainan kongenital.Kesimpulan: 0.1µl Gama Sleeved cryoloop merupakan pilihan untuk digunakan sebagai alat vitrifikasi blastokis. Data awal yang kami sampaikan dan kelahiran bayi dari program tersebut memberikan harapan untuk kesuksesan program simpan beku embrio di klinik Permata Hati RSUP DR Sardjito Yogyakarta.Kata kunci: kriopreservasi, blastokis, vitrifikasi ABSTRACTBackground: Vitrification has been applied succesfully in human embryo using grid, straw and cryoloop. Gama Sleeved is a home made device develop at Permata Hati. We assessed the survival rate of human blastocyst vitrified in 0.1µl Gama Sleeved cryoloop as device.Method: Excess good grade human D5 embryos were vitrified, upon a detailed informed consent. Embryos were hold in handling media (GMOPS Plus; Vitrolife) for 1 minute; (7.5% EG (v/v); 7.5% DMSO (v/v)) for 2-3 minutes, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) for 30 seconds at room temperature before inserted in to the loops, then directly plunged into the liquid nitrogen. Prior to ET, embryos were warmed by two step technique in sucrose 0.25M for 2 min and 0.125M sucrosa for 3 min. Embryos were then cultured.Results: Total of 97 vitrified warmed human blastocyst (67 patients) were used and 91 (93.8%) were transferred. Non-transferred blastocyst (6.2%) has more than 50% lyse. The clinical pregnancy rate was 43.9%. The implantation rate was 37.4%. Currently, 20 deliveries of 23 babies born from vitrified blastocyst using 0.1µl Gama Sleeved cryoloop, and another 5 ongoing pregnancy. So far there was 1 early pregnancy loss, 2 miscarriages at 12 weeks pregnancy, and one infant died due to a congenital anomaly.Conclusion: 0.1µl Gama Sleeved cryoloop provides an excellent alternative to existing vitrification devices. These initial data and babies delivered from the program have been promising to a vitrification system in our own ART program.Keywords: cryopreservation, blastocyst, vitrification1Permata Hati Infertility Clinic RSUP DR Sardjito, Yogyakarta2Div Reproductive Endocrinology and Fertility OBGYN Medical Faculty Gadjah Mada University, Yogyakarta3Lab. Anatomi Embriologi FKH, Institut Teknologi Pertanian, Bogor4EPRD- Dept. Obstetrics and Gynecology, Monash University, Monash Medical Center,Victoria, Melbourne5Lab. Reproductive Physiology, Jenderal Soedirman University, Purwokerto Correspondence address: + 62 274 518684; fax + 62 274 553575; email: itafauzia@yahoo.com
MICROVOLUME OF 0.1µL GAMA SLEEVED CRYOLOOPS FOR BLASTOCYST VITRIFICATION OF ASSISTED REPRODUCTIVE TECHNOLOGY PATIENTS Hanoum, Ita Fauzia; Boediono, Arief; Pangestu, Mulyoto; Haryadi, Dwi; Widad, Shofwal; Dasuki, Djaswadi
JURNAL KESEHATAN REPRODUKSI Vol 2, No 1 (2015)
Publisher : IPAKESPRO

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (181.875 KB)

Abstract

Ita Fauzia Hanoum1,2, Arief Boediono3, Mulyoto Pangestu4,5, Dwi Haryadi1,Shofwal Widad1,2, Djaswadi Dasuki1,2 ABSTRAK Latar Belakang: Prosedur embrio vitrifikasi menggunakan alat berupa grid, straw atau cryoloop. Gama Sleeved cryoloop dibuat dan dikembangkan di klinik Permata Hati. Untuk itu, dilakukan pengamatan keberhasilan prosedur vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop.Metode: Vitrifikasi dilakukan pada blastokis dengan kualitas baik yang diperoleh pada hari ke 5 setelah fertilisasi. Inform consent telah disampaikan sebelumnya kepada pasien program bayi tabung di Klinik Permata Hati. Prosedur dilakukan dengan menggunakan media handling (GMOPS Plus; Vitrolife) embrio diinkubasi selama 1 menit; (7.5% EG (v/v); 7.5% DMSO (v/v)) selama 2-3 menit, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) selama 30 detik pada suhu ruang sebelum kemudian diletakkan di dalam cryoloop, setelah itu secara cepat cryoloop yang berisi embrio dibenamkan ke dalam nitrogen cair. Sebelum dilakukan embryo transfer (ET), embrio dihangatkan dengan cara two step technique (sucrose 0.25M) selama 2 menit dan selama 3 menit (sucrose 0.125M).Hasil: Sejumlah 97 blastokis divitrifikasi dan dihangatkan (67 pasien), dimana 91 blastokis berhasil ditransfer ke rahim ibu (93.8%). Blastokis yang tidak berhasil selamat dari prosedur penghangatan adalah blastokis dengan kerusakan lebih dari 50%. Diperoleh kehamilan klinis 43.3% sedangkan angka implantasi adalah 37.4%. Sampai saat ini, dilaporkan 20 kelahiran (23 bayi) dari program vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop, sementara 5 kehamilan masih berlangsung. Satu kehamilan dilaporkan gugur pada usia kehamilan yang masih sangat awal, dua keguguran pada usia kehamilan 12 minggu dan satu bayi lahir meninggal karena kelainan kongenital.Kesimpulan: 0.1µl Gama Sleeved cryoloop merupakan pilihan untuk digunakan sebagai alat vitrifikasi blastokis. Data awal yang kami sampaikan dan kelahiran bayi dari program tersebut memberikan harapan untuk kesuksesan program simpan beku embrio di klinik Permata Hati RSUP DR Sardjito Yogyakarta.Kata kunci: kriopreservasi, blastokis, vitrifikasi ABSTRACTBackground: Vitrification has been applied succesfully in human embryo using grid, straw and cryoloop. Gama Sleeved is a home made device develop at Permata Hati. We assessed the survival rate of human blastocyst vitrified in 0.1µl Gama Sleeved cryoloop as device.Method: Excess good grade human D5 embryos were vitrified, upon a detailed informed consent. Embryos were hold in handling media (GMOPS Plus; Vitrolife) for 1 minute; (7.5% EG (v/v); 7.5% DMSO (v/v)) for 2-3 minutes, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) for 30 seconds at room temperature before inserted in to the loops, then directly plunged into the liquid nitrogen. Prior to ET, embryos were warmed by two step technique in sucrose 0.25M for 2 min and 0.125M sucrosa for 3 min. Embryos were then cultured.Results: Total of 97 vitrified warmed human blastocyst (67 patients) were used and 91 (93.8%) were transferred. Non-transferred blastocyst (6.2%) has more than 50% lyse. The clinical pregnancy rate was 43.9%. The implantation rate was 37.4%. Currently, 20 deliveries of 23 babies born from vitrified blastocyst using 0.1µl Gama Sleeved cryoloop, and another 5 ongoing pregnancy. So far there was 1 early pregnancy loss, 2 miscarriages at 12 weeks pregnancy, and one infant died due to a congenital anomaly.Conclusion: 0.1µl Gama Sleeved cryoloop provides an excellent alternative to existing vitrification devices. These initial data and babies delivered from the program have been promising to a vitrification system in our own ART program.Keywords: cryopreservation, blastocyst, vitrification1Permata Hati Infertility Clinic RSUP DR Sardjito, Yogyakarta2Div Reproductive Endocrinology and Fertility OBGYN Medical Faculty Gadjah Mada University, Yogyakarta3Lab. Anatomi Embriologi FKH, Institut Teknologi Pertanian, Bogor4EPRD- Dept. Obstetrics and Gynecology, Monash University, Monash Medical Center,Victoria, Melbourne5Lab. Reproductive Physiology, Jenderal Soedirman University, Purwokerto Correspondence address: + 62 274 518684; fax + 62 274 553575; email: itafauzia@yahoo.com
JUMLAH SEL PIRAMIDAL CA3 HIPOKAMPUS TIKUS PUTIH JANTAN PADA BERBAGAI MODEL STRES KERJA KRONIK Arjadi, Fitrianto; Soejono, Sri Kadarsih; Maurits, Lientje Setyawati; Pangestu, Mulyoto
Majalah Kedokteran Bandung Vol 46, No 4 (2014)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (556.626 KB)

Abstract

Paparan stres kronik dan berkepanjangan mengakibatkan hilangnya neuron di regio  CA3 (cornu ammonis) hipokampus dan  penurunan kognitif. Tujuan penelitian  mengetahui perbedaan jumlah sel piramidal CA3 hipokampus tikus putih jantan pada model stres kerja meliputi   paradoxical sleep deprivation (PSD), imobilisasi, dan footshock kronik. Metode penelitian adalah  post-test only with control group design experimental dengan rancangan acak lengkap  menggunakan 24 ekor tikus putih jantan galur Wistar usia 3?4 bulan yang dibagi 4 kelompok:  KI kontrol tanpa perlakuan,  KII (stres PSD), KIII (stres imobilisasi), dan KIV (stres footshock) dan sel piramidal CA3 hipokampus  diwarnai dengan toluidine-blue. Jumlah sel dihitung menggunakan  perangkat lunak Image raster v2.1, perbesaran 400x tiap 10 lapangan pandang. Penelitian dilakukan  6 bulan (April?September 2012) di Laboratorium Hewan Coba, Fakultas Kedokteran dan Ilmu Kesehatan Universitas Jenderal Soedirman. Analisis data menggunakan analysis of variance (ANOVA) dengan Post-Hoc least significant difference (LSD) menunjukkan  perbedaan jumlah sel piramidal CA3 hipokampus signifikan (p=0,037) pada minimal dua kelompok perlakuan. Uji statistik dengan Post-Hoc LSD menunjukkan perbedaan signifikan jumlah sel piramidal CA3 hipokampus antara  kelompok kontrol (12,9±2,47) dan kelompok imobilisasi (9,00±1,53) (p<0,05). Simpulan, kelompok imobilisasi kronik memiliki jumlah sel piramidal CA3 hipokampus terendah dibandingkan dengan ketiga kelompok lainnya.Kata kunci:  Model stres kerja kronik, sel piramidal CA3 hipokampus, tikus putih jantanNumber of CA3 Pyramidal Cell in Male Albino Rat   Hippocampus Exposed to Various Chronic Work Stress Models AbstractProlonged and chronic exposure to stress leads to the loss of neurons at the CA3 (cornu ammonis) hippocampus region and spatial memory deficits. The aim of this study was to study the number of CA3 pyramidal cells in albino rats that were exposed to chronic stress of works model consisting of paradoxical sleep deprivation (PSD), immobilization, and foot shock stresses. The method applied was the post-test only method with control group experimental design using completed randomized design (CRD on 24 3?4 month old male Wistar rats . The rats were divided into 4 groups: group I (control), group II (PSD stress), group III (immobilization stress), and group IV (footshock stress). The CA3 pyramidal cell hippocampus was stained with toluidine-blue. The number of CA3 pyramidal cell of hippocampus was counted using Image raster v2.1 software at 400x magnification in 10 duplicates for each sample. The study was conducted in six months (April?September 2012) at the Animal Laboratory, Faculty of Medical and Health Sciences, Jenderal Soedirman University. Analysis for the differences in the number of CA3 pyramidal cells was conducted using analysis of variance (ANOVA) with Post-Hoc LSD. The results of the ANOVA  showed a p value=0.037, meaning that there was significant difference in at least two groupsof treatment. Further statistical test using Post-Hoc LSD showed a significant difference  between the control group (12.9 ± 2.47) and the chronic immobillization group (9,00 ± 1,53) (p<0.05). In conclusion, the chronic immobillization stress group has the lowest average number of hippocampus CA3 pyramidal cells compared to other groups Key words: CA3 pyramidal cell in hippocampus, chronic works stress model, male albino rats DOI: 10.15395/mkb.v46n4.337
SPERM PRESERVATION USING FREEZE-DRYING METHOD SAILI, TAKDIR; PANGESTU, MULYOTO; SETIADI, MOHAMAD AGUS; AGUNGPRIYONO, SRIHADI; R. TOELIHERE, MOZES; BOEDIONO, ARIEF
HAYATI Journal of Biosciences Vol. 12 No. 1 (2005): March 2005
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (32.025 KB) | DOI: 10.4308/hjb.12.1.41

Abstract

Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI) for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF) the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.
MICROVOLUME OF 0.1µL GAMA SLEEVED CRYOLOOPS FOR BLASTOCYST VITRIFICATION OF ASSISTED REPRODUCTIVE TECHNOLOGY PATIENTS Hanoum, Ita Fauzia; Boediono, Arief; Pangestu, Mulyoto; Haryadi, Dwi; Widad, Shofwal; Dasuki, Djaswadi
JURNAL KESEHATAN REPRODUKSI Vol 2, No 1 (2015)
Publisher : IPAKESPRO

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (181.875 KB) | DOI: 10.22146/jkr.7127

Abstract

Ita Fauzia Hanoum1,2, Arief Boediono3, Mulyoto Pangestu4,5, Dwi Haryadi1,Shofwal Widad1,2, Djaswadi Dasuki1,2 ABSTRAK Latar Belakang: Prosedur embrio vitrifikasi menggunakan alat berupa grid, straw atau cryoloop. Gama Sleeved cryoloop dibuat dan dikembangkan di klinik Permata Hati. Untuk itu, dilakukan pengamatan keberhasilan prosedur vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop.Metode: Vitrifikasi dilakukan pada blastokis dengan kualitas baik yang diperoleh pada hari ke 5 setelah fertilisasi. Inform consent telah disampaikan sebelumnya kepada pasien program bayi tabung di Klinik Permata Hati. Prosedur dilakukan dengan menggunakan media handling (GMOPS Plus; Vitrolife) embrio diinkubasi selama 1 menit; (7.5% EG (v/v); 7.5% DMSO (v/v)) selama 2-3 menit, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) selama 30 detik pada suhu ruang sebelum kemudian diletakkan di dalam cryoloop, setelah itu secara cepat cryoloop yang berisi embrio dibenamkan ke dalam nitrogen cair. Sebelum dilakukan embryo transfer (ET), embrio dihangatkan dengan cara two step technique (sucrose 0.25M) selama 2 menit dan selama 3 menit (sucrose 0.125M).Hasil: Sejumlah 97 blastokis divitrifikasi dan dihangatkan (67 pasien), dimana 91 blastokis berhasil ditransfer ke rahim ibu (93.8%). Blastokis yang tidak berhasil selamat dari prosedur penghangatan adalah blastokis dengan kerusakan lebih dari 50%. Diperoleh kehamilan klinis 43.3% sedangkan angka implantasi adalah 37.4%. Sampai saat ini, dilaporkan 20 kelahiran (23 bayi) dari program vitrifikasi menggunakan 0.1µl Gama Sleeved cryoloop, sementara 5 kehamilan masih berlangsung. Satu kehamilan dilaporkan gugur pada usia kehamilan yang masih sangat awal, dua keguguran pada usia kehamilan 12 minggu dan satu bayi lahir meninggal karena kelainan kongenital.Kesimpulan: 0.1µl Gama Sleeved cryoloop merupakan pilihan untuk digunakan sebagai alat vitrifikasi blastokis. Data awal yang kami sampaikan dan kelahiran bayi dari program tersebut memberikan harapan untuk kesuksesan program simpan beku embrio di klinik Permata Hati RSUP DR Sardjito Yogyakarta.Kata kunci: kriopreservasi, blastokis, vitrifikasi ABSTRACTBackground: Vitrification has been applied succesfully in human embryo using grid, straw and cryoloop. Gama Sleeved is a home made device develop at Permata Hati. We assessed the survival rate of human blastocyst vitrified in 0.1µl Gama Sleeved cryoloop as device.Method: Excess good grade human D5 embryos were vitrified, upon a detailed informed consent. Embryos were hold in handling media (GMOPS Plus; Vitrolife) for 1 minute; (7.5% EG (v/v); 7.5% DMSO (v/v)) for 2-3 minutes, (15% EG (v/v); 15% DMSO v/v; 10 mg/ml Ficoll; 0.65 M Sucrosa) for 30 seconds at room temperature before inserted in to the loops, then directly plunged into the liquid nitrogen. Prior to ET, embryos were warmed by two step technique in sucrose 0.25M for 2 min and 0.125M sucrosa for 3 min. Embryos were then cultured.Results: Total of 97 vitrified warmed human blastocyst (67 patients) were used and 91 (93.8%) were transferred. Non-transferred blastocyst (6.2%) has more than 50% lyse. The clinical pregnancy rate was 43.9%. The implantation rate was 37.4%. Currently, 20 deliveries of 23 babies born from vitrified blastocyst using 0.1µl Gama Sleeved cryoloop, and another 5 ongoing pregnancy. So far there was 1 early pregnancy loss, 2 miscarriages at 12 weeks pregnancy, and one infant died due to a congenital anomaly.Conclusion: 0.1µl Gama Sleeved cryoloop provides an excellent alternative to existing vitrification devices. These initial data and babies delivered from the program have been promising to a vitrification system in our own ART program.Keywords: cryopreservation, blastocyst, vitrification1Permata Hati Infertility Clinic RSUP DR Sardjito, Yogyakarta2Div Reproductive Endocrinology and Fertility OBGYN Medical Faculty Gadjah Mada University, Yogyakarta3Lab. Anatomi Embriologi FKH, Institut Teknologi Pertanian, Bogor4EPRD- Dept. Obstetrics and Gynecology, Monash University, Monash Medical Center,Victoria, Melbourne5Lab. Reproductive Physiology, Jenderal Soedirman University, Purwokerto Correspondence address: + 62 274 518684; fax + 62 274 553575; email: itafauzia@yahoo.com