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Hasil Pencarian

Ditemukan 5 dokumen yang sesuai dengan query
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Adrian Pragiwaksana
Abstrak :
Sel punca mesenkim (MSC) dan sel punca pluripoten terinduksi (iPSC) telah dilaporkan mampu berdiferensiasi menjadi hepatosit secara in vitro dengan berbagai tingkat maturasi hepatosit. Sebuah metode sederhana untuk proses deselulerisasi perancah hati telah dikembangkan oleh Fakultas Kedokteran Universitas Indonesia. Penelitian ini bertujuan untuk mengevaluasi diferensiasi hepatosit dari iPSC dibandingkan dengan MSC dalam perancah hati yang dideselularisasi. Langkah pada penelitian ini adalah mengkultur iPSC dan MSC, mendeselularisasi hati kelinci, menyemai kultur sel ke dalam perancah, dan mendiferensiasikan menjadi hepatosit selama 21 hari dengan protokol Blackford yang dimodifikasi. Pemeriksaan dilakukan dengan pewarnaan Haematoxylin Eosin (HE), Masson Trichrome (MT), imunohistokimia (IHK) albumin dan cytochrome 3A4 (CYP3A4). Ekspresi gen albumin, cytochrome P450 (CYP450), dan cytokeratin-19 (CK-19) dianalisis menggunakan qRT-PCR. Pemeriksaan scanning electron microscope (SEM) dan immunofluorescence (IF) marker hepatocyte nuclear factor 4 alpha (HNF4-α) dan CCAAT/enhancer-binding protein alpha (CEBPA) dilakukan. Diferensiasi hepatosit dari iPSC dalam perancah hati yang dideselulerisasi dibandingkan dengan diferensiasi hepatosit dari MSC dalam perancah hati yang dideselulerisasi menunjukkan pembentukan sel tunggal dan kapasitas adhesi pada perancah yang lebih sedikit, dan penurunan tren ekspresi albumin dan CYP450 yang lebih rendah. Jumlah penyemaian sel awal yang lebih rendah menyebabkan hanya beberapa iPSC menempel pada bagian-bagian tertentu dari perancah hati yang dideselularisasi. Injeksi jarum suntik manual untuk reselulerisasi yang tidak merata menciptakan pola pembentukan sel tunggal oleh hepatosit dari diferensiasi iPSC di perancah hati yang dideselulerisasi. Hepatosit dari diferensiasi MSC memiliki kapasitas adhesi lebih tinggi ke perancah hati yang dideselulerisasi yang mengarah pada peningkatan tren ekspresi albumin dan CYP450. Penurunan ekspresi gen CK-19 lebih banyak terjadi pada diferensiasi hepatosit dari iPSC. Hasil tersebut dikonfirmasi oleh adanya sinyal positif protein HNF4-α dan CEBPA dengan pemeriksaan IF yang menunjukkan hepatosit yang dewasa. Kesimpulan dari penelitian ini adalah diferensiasi hepatosit dari iPSC pada perancah hati yang dideselularisasi lebih dewasa dengan adhesi sel-matriks ekstraseluler lebih rendah, distribusi sel spasial saling berjauhan, dan ekspresi albumin dan CYP450 lebih rendah dibandingkan dengan diferensiasi hepatosit dari MSC pada perancah hati yang dideselularisasi. ......Mesenchymal stem cells (MSC) and induced pluripotent stem cells (iPSC) have been reported able to differentiate to hepatocyte in vitro with varying degree of hepatocyte maturation. A simple method to decellularized liver scaffold has been established by Faculty of medicine Universitas Indonesia. This study aims to evaluate hepatocyte differentiation from iPSCs compared to MSCs in decellularized liver scaffold. iPSCs and MSCs were cultured, rabbit liver were decellularized, cell cultures were seeded into the scaffold, and differentiated into hepatocytes for 21 days with modified Blackford protocol. Haematoxylin-Eosin (HE), Masson Trichrome (MT), immunohistochemistry (IHC) albumin and CYP3A4 was performed. Expression of albumin, cytochrome P450 (CYP450) and cytokeratin-19 (CK-19) genes were analyzed using qRT-PCR. Scanning electron microscope (SEM) and immunofluorescence (IF) examination of hepatocyte nuclear factor 4 alpha (HNF4-α) and CCAAT/enhancer-binding protein alpha (CEBPA) marker was performed. Hepatocyte differentiated iPSCs compared with hepatocyte differentiated MSCs in decellularized liver scaffold single–cell–formation and lower adhesion capacity in scaffold, and decrease trends of albumin and CYP450 expression. Lower initial seeding cell number causes only a few iPSCs to attach to certain parts of decellularized liver scaffold. Manual syringe injection for recellularization abruptly and unevenly create pattern of single–cell–formation by hepatocyte differentiated iPSCs in the decellularized liver scaffold. Hepatocyte differentiated MSCs have higher adhesion capacity to decellularized liver scaffold that lead to increase trends of albumin and CYP450 expression. CK-19 expression gene diminished more prominent in hepatocyte differentiated iPSCs. These results were confirmed by the presence of HNF4-α and CEBPA positive signal protein with IF examination, showing mature hepatocyte.The conclusion of this study is hepatocyte differentiated iPSCs in decellularized liver scaffold differentiation is more mature with lower cell-extracelullar matrix adhesion, spatial cell distribution far from each other, and lower albumin and CYP450 expression than hepatocyte differentiated MSCs in decellularized liver scaffold.
Depok: Fakultas Kedokteran Univesitas Indonesia, 2021
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UI - Tesis Membership  Universitas Indonesia Library
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Abstrak :
This volume looks at induced pluripotent stem (iPS) cells, mature cells that have been genetically reprogrammed so that they return to their embryonic state.
New York: Springer, 2012
e20401549
eBooks  Universitas Indonesia Library
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Amit, Michal, editor
Abstrak :
Human pluripotent stem cells, including human embryonic stem cells and induced pluripotent stem cells, are a key focus of current biomedical research. The emergence of state of the art culturing techniques is promoting the realization of the full potential of pluripotent stem cells in basic and translational research and in cell-based therapies. This comprehensive and authoritative atlas summarizes more than a decade of experience accumulated by a leading research team in this field. Hands-on step-by-step guidance for the derivation and culturing of human pluripotent stem cells in defined conditions (animal product-free, serum-free, feeder-free) and in non-adhesion suspension culture are provided, as well as methods for examining pluripotency (embryoid body and teratoma formation) and karyotype stability.
New York: Springer, 2012
e20401870
eBooks  Universitas Indonesia Library
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Vivi M. Heine
Abstrak :
This book discusses the various methods to reprogram cells, the control and determination of cell identity, the epigenetic models that have emerged and the application of iPS cell therapy for brain diseases, in particular Parkinson’s disease and Vanishing White Matter (VWM).​
Dordrecht: [, Springer], 2012
e20417567
eBooks  Universitas Indonesia Library
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Lee, Yee-Ki
Abstrak :
Calcium is crucial in governing contractile activities of myofilaments in cardiomyocytes, any defeats in calcium homeostasis of the cells would adversely affect heart pumping action. The characterization of calcium handling properties in human induced pluripotent stem cell-derived cardiomyocytes (iPS-CMCs) is of significant interest and pertinent to the stem cell and cardiac regenerative field because of their potential patient-specific therapeutic use.
New York: Springer, 2012
e20401427
eBooks  Universitas Indonesia Library