Repository Universitas Pakuan

Detail Karya Ilmiah Dosen

Siti Warnasih,Winda Yulia, Benediktus Yohan, I Made Artika, R. Tedjo Sasmono

Judul : ISOLASI PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) DARI DARAH MANUSIA SEHAT DENGAN METODE SENTRIFUGASI GRADIEN FICOLL

Abstrak :
Peripheral blood sel mononuclear cell (PBMC) is a white blood cell that can be isolated using the gradient centrifugation method. Ficoll is a reactant which can be used in this method because it produce a high viability i.e. 97.1% ± 1.0%. This research aims to isolate PBMC from healthy human blood. PBMC’s result is used for analysis of dengue virus immune response.Dengue viruses infect PBMC, so going on the secretion of cytokine which is one marker of severity of dengue disease. Expression of cytokines can be measured using quantitative real-time reverse transcription – polymerase chain reaction (qRT-PCR).Venous blood from healthy human is takenaseptically using anticoagulants heparin, then PBMC is isolated with ficoll gradient centrifugation method to produce buffy coat that contain PBMC. Viability of PBMC is calculated using hemositometer with the help of a microscope.Based on the results obtained by PBMC isolation per mL of venous blood that is 2, 08x106 up to 2, 61x106 with an average of 2, i.e. 31x106 withviabilityof 95-99%. These results indicate that this method is quite effective in isolating PBMC from healthy human blood.

Tahun : 2015 Media Publikasi : Jurnal Nasional Blm Akreditasi

Kategori : Jurnal No/Vol/Tahun : 1 / 16 / 2016

ISSN/ISBN : 1411-9447

PTN/S : Universitas Pakuan Program Studi : KIMIA

Bibliography :
Abbas AK, Lichtman AH, Pillai S. 2012. Cellularand Molecular Immu nology. 7thed. United States of America: Elsevier.

Almeida, M.C., Silva, A.C., Barral, A., Netto, M.B. 2000. A simple Method for human peripheral blood monocyte isolation. Mem Inst Oswaldo Cruz. 95(2):221-223.

Bahunde, F., Awoyode, R., Fields, B., McLean, P., Tambwe, C., Johnson, N. 2013. Creating Evidence-based Procedures Out of Establised Processes: Validation of Ficoll PlaqueTM Centrifugation for Isolation of Peripheral Blood Mononuclear Cells. In Precision Bioservices, Inc. Frederick. MD Viability and Purity of Cell.

Beccera, A., Warke, R.V., Martin, K., Xhaja, K., de Bosch, N., Rothman, A.L., Bosch, I. 2009. Gene expression profiling of dengue infected human primary cells identifies secreted mediators in vivo. J Med Virol. 81(8):1403-1411.

Boyum, A., Scand.1976. Isolation of lymphocytes, granulocytes and macrophages. J ImmunolSuppl. 5: 9–15.

Chang, Y., Hsieh, P-H., Chao, C.C-K. 2009. The efficiency of percoll and ficoll density gradient media inthe isolation of marrow derived human mesenchymal stemcells with osteogenic potential. Med J. 32(3): 264-274.

Delves, P.J., Martin, S.J., Burton, D.R., Roitt, I.M. 2006. Roitt’s Essential Immunology. 11th ed. Blackwell Publishing. Graham JM. 2002. Isolation of peripheral blood mononuclear cells from macaques on a density barrier. The Scientific World Journal 2:1654– 1656.

Gubler, D.J. 1998. Dengue and dengue hemorrhagic fever. Clinical Microbiology Reviews. 11(3):480498.

Healthcare GE. 2014. Isolation of mononuclear cells; Methodology and applications. GE Healthcare.

Kou, Z., Quinn, M., Chen, H., Rodrigo, W.W., Rose, R.C., Schlesinger, J.J., Jin, X. 2008. Monocytes, but not T or B cells, are the principal target cells for dengue virus (DV) infection among human peripheral blood mono nuclear cells. J Med. Virol. 80: 134-146.

Nilsson, C.,Aboud, S., Karle´n, K., Hejdeman, B., Urassa, W., Biberfeld, G. 2008. Optimal Blood Mononuclear Cell Isolation Procedures for Gamma Interferon Enzyme-Linked Immuno spot Testing of Healthy Swedish and Tanzanian Subjects. Clinical and Vaccine Immunology. 15(4):585589.

Noisakran, S., Onlamoon, N., Song prakhon, P., Hsiao, H-M., Choke phaibulkit, K., Perng, G.C. 2010. Cells in dengue virus infection in vivo. Adv Virol. Article ID 164878, 15 pages.

Sun, P., Kochel, T.J. 2013. The Battle between Infection and Host Immune Responses of Dengue Virus and Its Implication in Dengue Disease Pathogenesis. Sci World J.

Warke, R.V., Xhaja, K., Martin, K.J., Fournier, M.F., Shaw, S.K., Brizuela, N., et al. 2003. Dengue virus induces novel change in gene expression of human umbilical vein endothelial cells. J Virol. 7(21): 11822-11832.

Yuwono, T. 2010. Biologi Molekuler. Jakarta: Erlangga

URL : http://journal.unpak.ac.id/index.php/ekologia/article/view/58/37

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Detail Karya Ilmiah Dosen

Siti Warnasih,Winda Yulia, Benediktus Yohan, I Made Artika, R. Tedjo Sasmono

Judul	:	ISOLASI PERIPHERAL BLOOD MONONUCLEAR CELLS (PBMCS) DARI DARAH MANUSIA SEHAT DENGAN METODE SENTRIFUGASI GRADIEN FICOLL
Abstrak	:	Peripheral blood sel mononuclear cell (PBMC) is a white blood cell that can be isolated using the gradient centrifugation method. Ficoll is a reactant which can be used in this method because it produce a high viability i.e. 97.1% ± 1.0%. This research aims to isolate PBMC from healthy human blood. PBMC’s result is used for analysis of dengue virus immune response.Dengue viruses infect PBMC, so going on the secretion of cytokine which is one marker of severity of dengue disease. Expression of cytokines can be measured using quantitative real-time reverse transcription – polymerase chain reaction (qRT-PCR).Venous blood from healthy human is takenaseptically using anticoagulants heparin, then PBMC is isolated with ficoll gradient centrifugation method to produce buffy coat that contain PBMC. Viability of PBMC is calculated using hemositometer with the help of a microscope.Based on the results obtained by PBMC isolation per mL of venous blood that is 2, 08x106 up to 2, 61x106 with an average of 2, i.e. 31x106 withviabilityof 95-99%. These results indicate that this method is quite effective in isolating PBMC from healthy human blood.
Tahun	:	2015	Media Publikasi	:	Jurnal Nasional Blm Akreditasi
Kategori	:	Jurnal	No/Vol/Tahun	:	1 / 16 / 2016
ISSN/ISBN	:	1411-9447
PTN/S	:	Universitas Pakuan	Program Studi	:	KIMIA
Bibliography	:	Abbas AK, Lichtman AH, Pillai S. 2012. Cellularand Molecular Immu nology. 7thed. United States of America: Elsevier. Almeida, M.C., Silva, A.C., Barral, A., Netto, M.B. 2000. A simple Method for human peripheral blood monocyte isolation. Mem Inst Oswaldo Cruz. 95(2):221-223. Bahunde, F., Awoyode, R., Fields, B., McLean, P., Tambwe, C., Johnson, N. 2013. Creating Evidence-based Procedures Out of Establised Processes: Validation of Ficoll PlaqueTM Centrifugation for Isolation of Peripheral Blood Mononuclear Cells. In Precision Bioservices, Inc. Frederick. MD Viability and Purity of Cell. Beccera, A., Warke, R.V., Martin, K., Xhaja, K., de Bosch, N., Rothman, A.L., Bosch, I. 2009. Gene expression profiling of dengue infected human primary cells identifies secreted mediators in vivo. J Med Virol. 81(8):1403-1411. Boyum, A., Scand.1976. Isolation of lymphocytes, granulocytes and macrophages. J ImmunolSuppl. 5: 9–15. Chang, Y., Hsieh, P-H., Chao, C.C-K. 2009. The efficiency of percoll and ficoll density gradient media inthe isolation of marrow derived human mesenchymal stemcells with osteogenic potential. Med J. 32(3): 264-274. Delves, P.J., Martin, S.J., Burton, D.R., Roitt, I.M. 2006. Roitt’s Essential Immunology. 11th ed. Blackwell Publishing. Graham JM. 2002. Isolation of peripheral blood mononuclear cells from macaques on a density barrier. The Scientific World Journal 2:1654– 1656. Gubler, D.J. 1998. Dengue and dengue hemorrhagic fever. Clinical Microbiology Reviews. 11(3):480498. Healthcare GE. 2014. Isolation of mononuclear cells; Methodology and applications. GE Healthcare. Kou, Z., Quinn, M., Chen, H., Rodrigo, W.W., Rose, R.C., Schlesinger, J.J., Jin, X. 2008. Monocytes, but not T or B cells, are the principal target cells for dengue virus (DV) infection among human peripheral blood mono nuclear cells. J Med. Virol. 80: 134-146. Nilsson, C.,Aboud, S., Karle´n, K., Hejdeman, B., Urassa, W., Biberfeld, G. 2008. Optimal Blood Mononuclear Cell Isolation Procedures for Gamma Interferon Enzyme-Linked Immuno spot Testing of Healthy Swedish and Tanzanian Subjects. Clinical and Vaccine Immunology. 15(4):585589. Noisakran, S., Onlamoon, N., Song prakhon, P., Hsiao, H-M., Choke phaibulkit, K., Perng, G.C. 2010. Cells in dengue virus infection in vivo. Adv Virol. Article ID 164878, 15 pages. Sun, P., Kochel, T.J. 2013. The Battle between Infection and Host Immune Responses of Dengue Virus and Its Implication in Dengue Disease Pathogenesis. Sci World J. Warke, R.V., Xhaja, K., Martin, K.J., Fournier, M.F., Shaw, S.K., Brizuela, N., et al. 2003. Dengue virus induces novel change in gene expression of human umbilical vein endothelial cells. J Virol. 7(21): 11822-11832. Yuwono, T. 2010. Biologi Molekuler. Jakarta: Erlangga
URL	:	http://journal.unpak.ac.id/index.php/ekologia/article/view/58/37