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All Journal BALI MEDICAL JOURNAL (BMJ) journal of internal medicine International Journal of Biosciences and Biotechnology Jurnal Ilmu dan Kesehatan Hewan Buletin Veteriner Udayana Buletin Udayana Mengabdi INDONESIAN JOURNAL OF BIOMEDICAL SCIENCES Jurnal Veteriner Jurnal Biologi Udayana Microbiology Indonesia Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Kesmas: Jurnal Kesehatan Masyarakat Nasional ISM (Intisari Sains Medis) : Jurnal Kedokteran Journal of Global Pharma Technology
I NYOMAN MANTIK ASTAWA
Laboratorium Virologi, Departemen Penyakit Hewan, Fakultas Kedokteran Hewan, Universitas Udayana, Bali-Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Education Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology Public Health Veterinary

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Journal : International Journal of Biosciences and Biotechnology

 1 
CD4 COUNT FROM CRYOPRESERVATION OF BUFFY COAT AND PBMC Rasmaya Niruri; Inna Narayani; Wayan Tunas Artama; Mantik Astawa; Ahmad Hamim Sadewa
International Journal of Biosciences and Biotechnology Vol 2 No 2 (2015)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Abstract

This study aimed to determine CD4 count from cryopreservation of Buffy coat (BC) and PeripheralBlood Mononuclear Cell (PBMC) with and without ficoll. Fifteen EDTA Blood sample (2 ml for eachtube) were drawn from one adult healthy subject. The samples were categorized into five group beforemeasuring the CD4 level (which were fresh whole blood [Group(G)-I], BC without ficoll [fresh <GII>and frozen <G-III>] , and PBMC resulted from BC and ficoll isolation [fresh <G-IV> andfrozen <G-V>]. Each group was replicated three times. Blood storage before preparation was less thanfour hours. Two months cryopreservation using liquid nitrogen (in 40% FBS, 10% DMSO, and RPMI)was conducted. The mean value of CD4 count (cell /mu1) were 522 (G-I), 1410 (G-II), 906 (G-III), 807(G-IV), and 733 (G-V). CD4 count, after 2 month preservation in liquid nitrogen, of the BC sample (G-III) was higher (906 cell /mu1) than PBMC (G-IV) sample (733 cell /mu1).
EFFECT OF RESTING TIME ON PERIPHERAL BLOOD MONONUCLEAR CELL YIELDS Inna Narayani; Rasmaya Niruri; Nyoman Mantik Astawa
International Journal of Biosciences and Biotechnology Vol 3 No 2 (2016)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

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Abstract

Cryopreservation of PBMC (peripheral blood mononuclear cells) was done to preserve and analyze the number of PBMC derived from blood samples which come in at different time. The batch analysis wasperformed at the same time in order to reduce variations in results. The analysis on the cells numbers carried out after 1, 3, 6, 12, and 24 hours. Heparinized whole blood was collected from healthy subject by venipuncture, and stored at room temperature. Blood is processed by centrifugation in Ficoll density gradient following the established method of Balai Besar Veteriner Denpasar. Buffy coat layer was collected and washed twice with HBSS (Hank's balanced salt solution) and was counted in Turk’s solution. The cells were then dissolved in 1 ml of cold freezing medium containing 10% DMSO and 50% FBS (fetal bovine serum) and stored overnight at -80°C before storage in liquid nitrogen vessel for few weeks. The samples rapidly thawed in a water bath at 37°C and washed twice with PBS (phosphate buffered saline). The cells were stored in 4°C PBS and counted in Turk’s solution after 1, 3, 6, 12, and 24 hours. The results obtained were varied with a declining trend.
Co-Authors A. A. G. P. Wiraguna A.A.G. Sudewa AAG Budhitresna AAG Putra Ahmad Hamim Sadewa Aida Lousie Tenden Rompis Alberto Agustinho Pereira Da Costa Joao Anak Agung Ayu Mirah Adi Anak Agung Bagus Bramardipa Anak Agung Gede Sudewa Djelantik Anak Agung Keswari Krisnandika Anak Agung Ngurah Subawa Anak Agung Oka Wijaya Anak Agung Sagung Kendran and R. Kusnandi Andika Budi Kurnianto Anwar Santoso Arthawan Arthawan Bayu Setiabudi Berata , I Ketut Chandra Yowani Dewa Ayu Agus Sri Laksmi DWI SURYANTO Faiziah - G.A.M.K. Dewi Gusti Ayu Mayani Kristina Dewi Gusti Ayu Yuniati Kencana Gusti Ngurah Narendra Putra HARTANINGSIH - Hartaningsih . I D. N. Wibawa I Dewa Made Sukrama I Gede Ngurah Harry Wijaya Surya I GEDE PUTU WIRAWAN I Gusti Agung Arta Putra I Gusti Agung Ayu Suartini I Gusti Agung Dewi Sarihati I Gusti Agung Trisna Windiani I Gusti Ayu Putu Eka Pratiwi I Gusti Made Krisna Erawan I Gusti Ngurah Kade Mahardika I Gusti Ngurah Sudisma I K. Sukardika I Kadek Swastika I Ketut Berata I Ketut Eli Supartika I Ketut Junitha I Ketut Suada I Ketut Suada I Ketut Suastika I Ketut Suastika I KETUT SUATA I Ketut Suatha I Ketut Suwiyoga I Made Damriyasa I Made Dwinata I Made Galih Diparayoga I Made Kardena I Made Subrata I Made Sudarmaja I Made Sukada i Nengah Wandia I Nyoman Agus Bagiada I Nyoman Polos I Nyoman Suarsana I Nyoman Suartha I W. Wita, I W. I Wayan Bebas I Wayan Gorda I Wayan Masa Tenaya I Wayan Megadhana I Wayan Putu Sutirta Yasa I Wayan Suardana I Wayan Wita I.A.P. Apsari I.B.K. Suardana I.H. Utama I.W. Batan Ida Ayu Pasti Apsari Ida Bagus Kade Suardana Ida Bagus Made Oka Ida Bagus Ngurah Swacita Ida Bagus Oka Winaya Ida Bagus Suardana Ignatius Ferdi Yuatmadja Inna Narayani K. Sri Marhaeni Julyasih K. Suata K. Sukardika Kadek Karang Agustina Ketut Budiasa Ketut Santhia Adhy Putra Ketut Suata Luh Dewi Anggreni LUH PUTU AGUSTINI Luh Putu Agustini Luh Putu Wrasiati M.D. Rudyanto M.Pd S.T. S.Pd. I Gde Wawan Sudatha . Marissa Divia Dayanti Mudinillah, Adam N. T. Suryadhi Ngakan Putu Anom Harjana Ni Luh Putu Eka Diarthini NI LUH PUTU MANIK WIDIYANTI Ni Made Krisna Dewi Ni Made Suaniti NINING HARTANINGSIH Nyoman Agus Bagiada Nyoman Tigeh Suryadi Oka Lely Palagan Senopati Sewoyo Putu Ayu Asri Damayanti Putu Ayu Sisyawati Putriningsih Putu Sudiarta Rasmaya Niruri Rasmaya Niruri S. Soetjiningsih, S. S. Sotjiningsih S.K. Widyastuti sang gede purnama Siti Maryam Soetjiningsih - Soetjiningsih . Sri Kayati Widyastuti Sri Wahjuni SUMARNO Sutjahjo Suherman, Sutjahjo Suwarno - T. Sari Nindia Tjok Gede Oka Pemayun, Tjok Gede Oka Tjokorda Gde Agung Suwardewa Wayan Tunas Artama Wimpie I Pangkahila Yasunobu Matsumoto Yosevangelika Hutabarat Yoshihiro Hayashi