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Optimization of Activation Methods for Mouse Oocytes Using Calcium-free CZB Medium, SrCl2, and Cytochalasin B in Vitro

Jurnal Kedokteran Maranatha Vol 8, No 2 (2009)
Publisher : Universitas Kristen Maranatha

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Abstract

Embryonic stem cells can be obtained by generating an embryo through fertilization; however, an embryo can also be generated asexually through parthenogenesis. This procedure will overcome the ethical issues regarding the use of embryos initially generated for reproductive purposes. The aim of this study was to obtain an optimized oocyte activation method through parthenogenesis by using mice oocytes as a model. Ten mM SrCl2 and 5 µg/ml Cytochalasin B (CB) in calcium-free Chatot Ziomek Bavister (CZB) were used as a medium for an in vitro activation of mouse oocytes. Treatment combinations for the oocyte activation methods were (A) activation in CZB & SrCl2 (prepared in stock) for two hours and in CZB & CB for four hours; (B) activation in CZB & SrCl2 (fresh medium) for two hours and in CZB & CB for four hours; and (C) activation in CZB & CB & SrCl2 (fresh medium) for six hours. The results show that the activation rate of  mouse oocytes  with  method C has  been  the best among all the protocols. This optimized protocol clearly provides a new insight in the generation of embryos for further use, particularly for producing embryonic stem cells.

Potensi Terapi Sel Punca dalam Dunia Kedokteran dan Permasalahannya

Jurnal Kedokteran Maranatha Vol 8, No 1 (2008)
Publisher : Universitas Kristen Maranatha

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Abstract

Stem cell  has become a main focus for not only reseacrhers but also society due to its potency in cell-based therapy. Regardless ethical issues surrounding the human embryonic stem cell, adult human stem cell became the alternative of choice for transplantation. In the an effort to minimize ethical problems of human embryonic stem cell transplantation, many breaktroughs have been conducted, like ANT (Altered Nuclear Transfer) and iPS (Induced Pluripotent Stem Cell). Development of stem cell technology  in producing testing model will assist a lot in potential drug testing, which might decrease potential side effect and numbers of human clinical trial.  

PENGARUH MOTIVASI TERHADAP KINERJA PEGAWAI DENGAN VARIABEL PEMEDIASI KEPUASAAN KERJA PADA PDAM KOTA MADIUN

JRMA | Jurnal Riset Manajemen dan Akuntansi Vol 1, No 1 (2013)
Publisher : JRMA | Jurnal Riset Manajemen dan Akuntansi

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Abstract

Penelitian ini bertujuan untuk menguji pengaruh motivasi terhadap kepuasaan kerja pegawai, pengaruh motivasi terhadap kinerja pegawai, pengaruh kepuasaan kerja terhadap kinerja pegawai, dan menguji kepuasaan kerja sebagai variabel pemediasi antara motivasi dan kinerja pegawai. Responden sebanyak 155 pegawai tetap di PDAM Kota Madiun. Analisis data menggunakan metode regresi. Temuan menunjukan motivasi berpengaruh signifikan pada kepuasaan kerja, motivasi tidak berpengaruh signifikan terhadap kinerja pegawai dan kepuasaan kerja berpengaruh signifikan terhadap kinerja pegawai. Temuan juga menunjukan bahwa kepuasaan kerja merupakan variabel pemediasi antara motivasi dengan kinerja pegawai.

Perkembangan Praimplantasi Embrio Mencit dengan Materi Genetik yang Berasal dari Parental, Maternal, dan Inti Sel Somatik (PRE-IMPLANTATION DEVELOPMENT OF MOUSE EMBRYO WITH GENETIC MATERIAL DERIVED FROM PARENTAL, MATERNAL AND SOMATIC CELL NUCLEUS)

Jurnal Veteriner Vol 15, No 1 (2014)
Publisher : Jurnal Veteriner

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Abstract

Cloned embryo and parthenogenetic embryo are a potential source of stem cells for regenerativemedicine. Stem cells derived from those embryos are expected to overcome the ethical issues to the use offertilization embryos for therapeutic purposes. The pre-implantation development is a critical step fordeveloping embryos reach the blastocyst stage. The objectives in vivo of this research are to produce mousecloned embryo, parthenogenetic embryo, and fertilized embryo and to study stages of  in vitro pre-implantation development culture. In vivo fertilized embryos, mouse oocytes, and cumulus cells were usedin this study. Treatment was performed on female mice superovulated with PMSG and hCG injections.Two-cell stage of in vivo fertilized embryos were collected on the second day post hCG injection. Clonedembryos were produced through Somatic Cell Nuclear Transfer (SCNT), which included enucleation, nucleartransfer and artificial activation. Parthenogenetic embryos were produced with artificial activationtechnique. The result of the research indicated that SCNT application was able to produce cloned embryos which could develop to blastocyst stage (3,2%). In addition, artificial activation of oocytes could produceparthenogenetic embryos which were able to develop up to the blastocyst stage (8,6%). In conclusion,efficiency level of parthenogenetic embryos that is able to reach the blastocyst stage was higher than in thecloned embryos. Fertilized embryos shows a better development and more efficient compared to in vitrocloned embryos and parthenogenetic embryos cultures.

PCS-16 Determaining Surgical Method by Meniscectomy Induction on Garut sheep (Ovis aries) for early stage of Osteoarthritis

Hemera Zoa Proceedings of the 20th FAVA & the 15th KIVNAS PDHI 2018
Publisher : Hemera Zoa

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Abstract

Osteoarthritis (OA) is the most common joint disease that cause of pain and disability by various factors such as advanced age, obesity, trauma, and arthritis disease. These factors affect by degeneration of the cartilage surface, leading to loss of matrix include proteoglycan osteophyte formation, subcondral and synovial membrane affected. In the healthy joint, meniscus, articular cartilage, subchondral bone, and synovial membrane provide support to the joint. The meniscus is an important load bearing structure and has nutritive as well as lubricating properties in the knee joint as well (Little et al. 2010).Animal models are research materials that can be used in studying potential pathogenesis and therapy in various diseases in humans. Sheep are commonly large animal model of OA because of the availability, ease of handling, and have a similarities with humans in size and structure of joint. In the development of science, sheep can be used as an animal model in studying the pathogenesis of diseases in human orthopedics studies such as joints, ligaments, and bones. Garut sheep is an Indonesian germplasm indigenous that has the structure, density, and size of joint anatomy that are similar in human joints rather than other small animals. This is the basis of the utilization of Garut sheep as an animal model in human orthopaedic. (Little et al. 2010; Gregory et al. 2012).The aim of this study was to identify and analyze the determining surgical method by meniscectomy induction on Garut sheep with 8 weeks post meniscectomy observation for early stage of OA.

Direct and Indirect Effect of TNFα and IFNγ Toward Apoptosis in Breast Cancer Cells

Molecular and Cellular Biomedical Sciences Vol 2, No 2 (2018)
Publisher : Cell and BioPharmaceutical Institute

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Abstract

Background: Breast cancer (BC) is the leading cause of death cancer in women. Cancer therapies using TNFα and IFNγ have been recently developed by direct effects and activation of immune responses. This study was performed to evaluate the effects of TNFα and IFNγ directly, and TNFα and IFNγ secreted by Conditioned Medium-human Wharton’s Jelly Mesenchymal Stem Cells (CM-hWJMSCs) toward apoptosis of BC cells (MCF7).Materials and Methods: BC cells were induced by TNFα and IFNγ in 175 and 350ng/mL, respectively. CM-hWJMSCs were produced by co-culture hWJMSCs and NK cells that secreted TNFα, IFNγ, perforin (Prf1), granzyme B (GzmB) for treating BC cells. The BC cells were treated with CM-hWJMSCs in 50%. The expression of apoptotic genes Bax, p53, and the antiapoptotic gene Bcl-2 were determined using RT-PCR.Results: TNFα and IFNγ at concentration of 350 ng/mL induced higher Bax expression compared to 175 ng/mL. TNFα and IFNγ 350 ng/mL, 175 ng/mL induced p53 expression, whilst TNFα and IFNγ at 350 ng/mL decreased Bcl-2 expression. Perf1, GzmB, TNFα and IFNγ-containing CM-hWJMSCs induced significantly apoptosis percentage, induced Bax expression, but did not effect p53, Bcl-2 expression.Conclusion: TNFα and IFNγ directly induce Bax, p53, decrease Bcl-2 gene expression. The Prf1, GzmB, TNFα, IFNγ-containing CM-hWJMSCs induce apoptosis and Bax expression.Keywords: breast cancer, Wharton’s Jelly mesenchymal stem cells, TNFα, IFNγ