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"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).​"

"This volume looks at induced pluripotent stem (iPS) cells, mature cells that have been genetically reprogrammed so that they return to their embryonic state."

"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.

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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."

"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. "

"Penelitian mengenai sel punca serta aplikasinya di bidang biomedis telah mengalami perkembangan yang pesat. Namun, penggunaan sel punca embrionik dalam penanganan medis menimbulkan kekhawatiran terkait bioetika penggunaan embrio manusia, penolakan sel punca oleh sistem imun tubuh pasien, dan risiko terbentuknya teratoma. Hal tersebut menimbulkan urgensi untuk membuat sel punca pluripoten dari sel somatik yang telah terdiferensiasi. Metode induced pluripotent stem cells dapat dimanfaatkan untuk membuat sel-sel punca dari sel-sel yang telah terdiferensiasi. vesikel ekstraseluler yang dihasilkan oleh sel-sel iPSC juga menjanjikan dalam inovasi aplikasi iPSC minim risiko. Untuk dapat mengaplikasikan vesikel ekstraseluler iPSC sebagai penanganan medis dengan dosis yang tepat, diperlukan uji sterilitas dan karakterisasi profil metabolit vesikel ekstraseluler dari iPSC. Tujuan dari penelitian ini adalah untuk mengetahui sterilitas sampel vesikel ekstraseluler iPSC SCTE FKUI dan mengetahui karakteristik profil metabolit dari sampel vesikel ekstraseluler iPSC yang didapatkan di SCTE FKUI. Metode penelitian yang digunakan mencakup uji sterilitas menggunakan Respiratory Flow Chip Kit dan analisis profil metabolit menggunakan Ultra-Performance Liquid Chromatography-Mass Spectrometry yang dilanjutkan dengan analisis menggunakan perangkat lunak MassLynx 4.1. Hasil menunjukkan bahwa sampel vesikel ekstraseluler iPSC koleksi FKUI steril terhadap 23 patogen infeksi saluran pernapasan akut, dan memiliki profil metabolit berupa 12 metabolit dengan karakteristik beragam.

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The research on stem cells and their applications in the biomedical field has been advancing, but the usage of embryonic cells had provoked controversies regarding the bioethics of extracting human embryonic tissues and poses a risk of rejection from patients’ immune system and the possibility of teratoma formation. Hence, the urgency to utilize a stem cell derived from terminally differentiated cells comes to surface. The iPSC method can be used to reprogram terminally differentiated cells back into cells with pluripotency. Extracellular vesicles secreted by iPSC are also promising for biomedical treatment. In order to use extracellular vesicles for biomedical treatment with a suitable dosage, it is necessary to confirm the sterility and characterize the extracellular vesicles of iPSC used. This research is aimed to test the sterility and analyze the metabolite profile of vesikel ekstraseluler iPSC samples from SCTE FKUI collection. Respiratory Flow Chip Kit is used as sterility test. Ultra-performance liquid chromatography-mass spectrometry is used to to analyze metabolite profile. The analysis of the metabolite profile is conducted with Masslynx 4.1. Result of the sterility test shows that the vesikel ekstraseluler iPSC collection was sterile from 23 pathogens of acute respiratory infections. Moreover, it is confirmed from LCMS analysis that the sample contains 12 metabolites with various characteristics.

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"The goal of this book is to provide an up-to-date review of the various types of blood-derived cells with regenerative capacity, identify opportunities for intervention by examining specific clinical applications, and recognize the regulatory environment that will encompass future therapies in regenerative medicine."

"Latar belakang: Hati kelinci yang dideselularisasi sebagai perancah untuk kultur organoid hati telah menunjukkan hasil yang menjanjikan dalam meningkatkan viabilitas dan fungsinya. Penelitian ini bertujuan untuk mengembangkan organoid dari kokultur sel yang dapat mendukung fungsi hati. Pemanfaaatan perancah hati yang dideselularisasi untuk mempertahankan viabilitas dan fungsionalitas hepatosit dan mengevaluasi organoid hati manusia yang ditransplantasikan ke model hewan coba dan mengetahui respons yang dimediasi sel imun. Metode: Sel hepatosit yang berasal dari iPSC manusia dikokultur dengan tiga sel lain untuk membentuk organoid hati. Delapan belas belas kelinci putih berusia 3 bulan digunakan dalam percobaan ini dan dibagi menjadi empat kelompok: Kelompok sham-operated (n = 3), kelompok ligasi duktus biliaris (n = 6), kelompok eksperimen dengan ligasi saluran empedu diikuti oleh transplantasi organoid hati (kelompok jangka pendek, (n=5); kelompok jangka panjang, (n=4).Hasil: Pada penelitian ini dilakukan analisis survival menggunakan metode Kaplan-Meier (KM) untuk menentukan probabilitas kumulatif kelangsungan hidup dari kejadian kematian pada kedua kelompok dengan dan tanpa transplantasi organoid hati. Hasil tes log-rank menunjukkan bahwa kemungkinan bertahan hidup secara keseluruhan antara kedua kelompok yang menerima perlakuan berbeda. (p=0,003). Kelompok jangka pendek menunjukkan peningkatan fungsi hati seperti albumin, CYP3A, dan tingkat AST yang lebih rendah daripada kelompok jangka panjang. Hati kelompok jangka pendek menunjukkan tingkat deposisi kolagen yang lebih rendah.Kesimpulan: Transplantasi organoid hati kokultur manusia dalam perancah hati yang dideselularisasi ke hewan yang diligasi duktus biliaris dapat mendukung kelangsungan hidup hewan dan fungsi hati untuk jangka pendek. Studi ini menyoroti potensi transplantasi organoid hati untuk mendukung fungsi hati jangka pendek. Namun, fungsi dan penolakan organoid hati dapat membatasi penggunaan pada jangka panjang.

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Background: Decellularized native liver scaffolds as a platform for liver organoid culture have shown promising results in improving their viability and function. This research aims to develop cocultured liver organoids that can recapitulate liver functions, utilize a decellularized native liver scaffold to maintain the viability and functionality of hepatocytes and evaluate human liver organoids transplanted into animal models to support liver function in two periods categories and immune-mediated response.

Methods: The hepatocyte-like cells derived from the human iPSCs were cocultured with three other cells to form liver organoids. Eighteen 3-month-old New Zealand White Rabbits were used in the experiment, divided into four groups: A sham-operated group (n=3), a bile duct ligation group (n=6), an experimental group with biliary duct ligation followed by liver organoid transplantation (short-term group, n=5; long-term group, n=4).

Results: We performed a survival analysis using the Kaplan-Meier (KM) method to determine the cumulative probability of survival from death events in both groups with and without liver organoid transplantation. The log-rank test results indicated a notable variation in the overall likelihood of survival between the two groups receiving different treatments. (p=0.003). The short-term group exhibited improved liver functions such as albumin, CYP3A, and lower levels of AST than the long-term group. The livers of the short-term group showed lower levels of collagen deposition.

Conclusions: Transplanting human coculture liver organoids in decellularized native liver scaffold into bile duct ligated animals could support the animal's survival and hepatic function for the short term. This study highlights the potential of liver organoid transplantation for short-term liver support. However, the functionality and rejection of liver organoids may limit their long-term use."

"This book reviews the current status of biomaterials for regenerative medicine, and highlights advances in both basic science and clinical practice. The latest methods for regulating the biological and chemical composition of biomaterials are described, together with techniques for modulating mechanical properties of engineered constructs. Contributors delineate methods for guiding the host response to implantable materials, and explain the use of biologically-inspired materials for optimal biological functionality and compatibility. "

"Chemical and functional genomic approaches to stem cell biology and regenerative medicine consolidates the latest information on functional genomics and chemical biology approaches that are used to study and control the fate of stem cells. It discusses new technologies and their recent applications in various areas of stem cell biology, covering :- The use of both embryonic and adult stem cells- A vast array of technologies, including: genome-wide expression analysis, functional genomic profiling with arrayed cDNA and RNAi expression libraries, informatics, chemical genomics, mass spectrometry, and proteomics- The applications of advanced technologies in various areas of stem cell biology, encompassing: self-renewal, differentiation, and reprogramming of different types of embryonic and adult stem cells, as well as regeneration in model organisms- Technological tools for studying the biology of stem cells"

"Human dental pulp of exfoliated deciduous teeth contains the population of cells that exhibited mesenchymal stem cell (MSC) characters. Though, a cell amplification process is indeed required to secure and adequate cell number for such a potential employment. Several publications suggested the alteration of MSCs upon in vitro culture, for example, the decrease in proliferation and the loss of stem cell characters. Here, we investigated an influence of basic fibroblast growth factor (bFGF) on stem cells isolated from human exfoliated deciduous teeth (SHEDs) with respect to cell proliferation, colony forming unit efficiency and stem cell marker expression in both short- and long-term cultures. For short-term bFGF treatment, SHEDs were treated with bFGF for 48h. While, in long-term bFGF supplementation, SHEDs were maintained in culture and continuous passage upon confluence in medium supplemented witg bFGF. Cells at passage (P) 5 and 10 were employed for characterization. Our result showed that short-term bFGF treatment enhanced OCT4, REX1, and NANOG mRNA expression as well as colony forming unit ability. The FGFR inhibitor pretreatment was able to attenuate the influence of bFGF on pluripotent stem cell marker expression, confirming bFGF function. In addition, cells cultured in high passage number had decreased in cell proliferation, colony forming unit capacity, and pluripotent stem cell marker mRNA expression. However, bFGF supplementation in culture medium enhanced both pluripotent stem cell marker expression and colony forming unit capacity in later passage, though the effect was not robust. Together, these results indicate that high passage number may attenuate pluripotent properties of SHEDs and bFGF supplementation could be the beneficial approach to maintain SHEDs' stemness properties. "