A liver organoid is an in vitro reconstruction of the liver that mimics the in vivo liver microstructure and performs liver functions. Liver organoids can be used for drug testing, as a model of liver disease pathogenesis, and as a bioartificial liver prototype material to develop promising alternative therapies for liver failure. In this study, we reconstructed liver organoids using primary rat hepatocytes, a hepatic stellate cell line (LX2), human umbilical cord-mesenchymal stem cells (UC-MSCs), and human umbilical cord blood (UCB)-CD34+ hematopoietic stem/progenitor cells. Suspensions of primary rat hepatocytes, LX2 cells, UC-MSCs, and UCB-CD34+ cells were co-cultured using 11 ratio sets, and spheroid formation was evaluated for 2 days. Ratio sets with a positive liver organoid appearance were cultured in four different culture media, and after they were harvested, their viability was compared with that of a hepatocyte monoculture. Liver organoids were further analyzed for 14 days to assess albumin and urea production as well as relative gene expression. We found that a 5:1:2:2 cellular density ratio of hepatocytes:LX2 cells:UC-MSCs:UCB-CD34+ cells, respectively, and Williams E medium supplemented with platelet lysate, ITS, and dexamethasone were the most suitable conditions for liver organoid reconstruction. Expression of the albumin and GPT1 genes and CD31 in the liver organoid increased until day 14, while urea secretion increased until day 5. Liver organoids reconstructed through the 3D co-culture of primary rat hepatocytes, LX2 cells, UC-MSCs, and UCB-CD34+ cells at a specific cellular ratio using an economical medium with a simple composition maintained their functions until day 14. As a future direction, these organoids can be used to develop a bioartificial liver.
Ascariasis is a global problem helath with transmission through soil contaminated withhelminth eggs. Personal and environmental hygiene as well as knowledge aboutascariasis affect the risk of ascariasis. The purpose of this community service is toincrease public knowledge about ascariasis. Outreach is given in poster mediapresentation, questions and answers with the community in Namorambe Village, DeliSerdang. Enthusiasm of the participants and the questions given showed the success ofthis activity. Education about personal and environmental hygiene and ascariasis neededto reduce the risk of ascariasis.
Background: Liver is a vital organ that has many functions including metabolism, detoxification of toxins and protein synthesis. The prevalence of liver disease is high and the treatment of liver disease continues to develop, so liver models are needed to study liver disease mechanisms and test drug toxicity. Many hepatocyte culture methods are being developed to construct an optimal liver model. The best culture method maintains high hepatocyte viability. Objective : This study aims to determine the viability of hepatocytes in 3D hanging drop method culture and in 2D culture. Methods: Hepatocytes were isolated from the liver of Sprague Dawley-Rats rats (n=2) and digested with Trypsin. Primary hepatocytes were cultured using the hanging drop method and conventional (2D) method. Hepatocyte viability was analyzed using the Trypan Blue Exclusion Test. Results: Hanging drop method had higher viability (81.18%) than the 2D culture method (69.22%) with p>0,05. Conclusion: The hanging drop method has better viability than the 2D culture method. Keywords: Hepatocyte, viability, 3D culture, 2D culture, hanging drop Latar belakang: Hati merupakan organ vital tubuh yang memiliki banyak fungsi diantaranya metabolisme, detoksifikasi racun dan sintesis protein. Prevalensi penyakit hati tinggi dan terapi penyakit hati terus berkembang, sehingga model hati sangat dibutuhkan untuk mempelahari mekanisme penyakit hati dan uji toksisitas obat. Banyak metode kultur hepatosit yang terus dikembangkan untuk menghasilkan suatu model hati yang optimal. Metode kultur yang terbaik adalah metode kultur dengan viabilitas hepatosit yang tinggi. Tujuan : Penelitian ini bertujuan untuk mengetahui viabilitas hepatosit pada kultur 3D metode hanging drop dan pada kultur 2D. Metode: Hepatosit diisolasi dari hati tikus Sprague Dawley-Rats (n=2) dan digesti dengan Tripsin. Hepatosit primer tikus dikultur dengan metode hanging drop (tetes gantung) dan metode konvensional (2D). Viabilitas hepatosit dianalisa dengan menggunakan Trypan Blue Exclusion Test. Hasil: Viabilitas hepatosit pada metode hanging drop memiliki viabilitas lebih tinggi (81,18%) dibandingan dengan metode kultur 2D (69,22%) dengan nilai p<5. Kesimpulan: Metode hanging drop memiliki viabilitas yang lebih baik dibandingkan metode kultur 2D. Kata Kunci: Hepatosit, viabilitas, kultur 3D, kultur 2D, hanging drop
Metabolic syndrome is a group of metabolic disorders with high prevalence in several populations and is a risk factor for cardiovascular disease and stroke with high mortality rate. Dyslipidemia in metabolic syndrome is marked by the increase of triglycerides. Apolipoprotein A5 (ApoA5) regulates triglycerides by lowering production of VLDL (Very Low Density Lipoprotein) and increases LPL (Lipoprotein Lipase) activity. Apolipoprotein A5 -1131T>C polymorphism in the promoter region is linked to the increase of triglycerides in metabolic syndrome. This study aims to find the relationship of apolipoprotein A5 -1131T>C polymorphism with triglycerides level in metabolic syndrome. This study was a case control study involving 50 subjects with 25 metabolic syndrome subjects and 25 nonmetabolic syndrome subjects. ApoA5 -1131T>C polymorphism was visualized with 5% agarose gel after restriction length fragment polymorphism (RFLP) digested with MseI. Data obtained were analyzed with unpaired t-test. Unpaired t-test result showed significant difference between ApoA5 -1131T>C polymorphism triglyceride level in metabolic syndrome and non-metabolic syndrome patient, be it TT or CC genotype with p<0.05. The TT genotype is found higher in metabolic syndrome compared to non-metabolic syndrome patients (72% vs 32%), CC genotype is found lesser in metabolic syndrome compared to non-metabolic syndrome (28% vs 68%), while heterozygous TC was not found in the subjects. This study showed that ApoA5 -1131T>C polymorphism was not a contributing factor for triglycerides level in metabolic syndrome. Increase in triglycerides in metabolic syndrome may be influenced by other ApoA5 SNPs.
The latest approach in tissue engineering techniques is the use of deselularized native organ as scaffolds for organ reconstruction. Scaffolds made from natural or synthetic matrix have many shortcomings in providing ideal extracellular matrix microstructure for cell proliferation and differentiation. Deselularized native scaffold can maintain the extracellular matrix of native organs, as an ideal microstructure scaffold for cell proliferation and differentiation. This technique answers the challenge of high need for liver transplants due to the limitations of liver donors and the need as liver models for drug testing.for drug testing. Liver is a complex organ, so ideal microstructure is needed in liver organoid reconstruction techniques to produce good synthetic and metabolic functions of the liver.
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