Using Microwell Chip for Scalable Generation of Pancreatic Aggregates
- سال انتشار: 1399
- محل انتشار: بیست و یکمین کنگره پزشکی تولید مثل و شانزدهمین کنگره زیست شناسی و فناوری سلول های بنیادی
- کد COI اختصاصی: RROYAN21_054
- زبان مقاله: انگلیسی
- تعداد مشاهده: 267
نویسندگان
Department of Genetics, Faculty of Basic Sciences and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran . Department of Stem Cells and Developmental Biology, Cell Science Research Center , Royan Institute for Stem Cell Biol
Department of Stem Cells and Developmental Biology, Cell Science Research Center , Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Department of Developmental Biology, Faculty of Basic Sciences and Advanced Technologies in Biolo
Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Department of Stem Cells and Developmental Biology, Cell Science Research Center , Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran,Department of Diabetes, Obesity and Metabolism, Cell Science Research Center, Royan Institute for
چکیده
Objective: For the production and bio-banking of organoids in large quantities for drug screening and cellular therapies, well-defined procedures for scalable production are required. As well, based on previous studies, co-culture with native pan-creatic mesenchyme promotes self-renewal and expansion of their epithelial progenitors. Therefore, in this study we used a microwell platform as a high throughput three-dimensional (۳D) culture system where human embryonic stem cells derived pancreatic progenitors (hESC-PPCs) can be co-cultured with human fetal pancreatic mesenchyme (hFP-MCs) for large-scale generation of pancreatic aggregates. The advantage of this plat-form compared to conventional suspension culture is its repro-ducibility.Materials and Methods: Non-adherent agarose microwell chip was fabricated by mold-replication technology which con-tained a structured polydimethylsiloxane (PDMS) surface with a standard array of ۴۰۰ µm diameter pyramidal microwells. The PPCs differentiation was achieved from Royan hESC lines. HFP-MCs were isolated from ۱۲-۲۰ week human embryos. For aggregate formation, cell mixture suspension of hESC-PPCs and hFP-MCs was seeded on microwell chips. After aggregates generated by force aggregation and ۲۴ hour incubation, were aspirated from the microwells and placed in static suspension culture.Results: This platform produces approximately ۴۷۰۰ aggre-gates where each aggregate has a diameter of ۷۰–۱۲۰ µm, lead-ing to uniformly sized, shaped and stable aggregates with low variation in diameter. We demonstrated that both cells are pre-sented inside the aggregates after ۴۸ hours of suspension cul-ture and formed aggregates can be harvested easily without any additional forces, which prevent damage to aggregates during harvesting. Aggregates were expressing PP- and MC specific gene and protein markers after ۴۸ hours culturing.Conclusion: Altogether, pancreatic aggregates which are pro-duced by our method formed round-shaped with well-defined borders aggregates and were able to be maintained stable in culture for short-range period in order to apply in in vivo trans-plantations and cell therapies.کلیدواژه ها
Microwell, Human Embryonic Stem Cell, Pancreatic Fetal Mesenchyme, Pancreatic Progenitor Cells, Organoidمقالات مرتبط جدید
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