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- Title
A Feasible and Efficient Protocol Developed for Scale up Differentiation of Human Pluripotent Stem Cells into Hepatocyte-Like Cells in A Single Suspension Culture.
- Authors
Vosough, M.; Omidinia, E.; Kadivar, M.; Shokrgozar, M. A.; Pournasr, B.; Aghdami, N.; Baharvand, H.
- Abstract
Objective: Recent papers described developing protocols to generate hepatocytes from pluripotent stem cells (PSCs) based on simulating developmental stages experimentally. Despite of these wonderful advances, due to inadequate amount of harvested cells their biomedical application is limited. In this regard, considering scale up cultures and practical differentiation protocols are two crucial factors. In this paper, we explain a single suspension culture and differentiation method for manufacturing functional Hepatocyte-Like Cells (HLCs) in stirrer bioreactor. Materials and Methods: Our stepwise protocol composed of serum starvation of cell aggregates, priming with rapamycin (Rapa) and direct induction of them with corresponding growth factors according to natural timetable in developmental biology. In comparison with other protocols in differentiation, this method is feasible, integrated and budget saving, giving the decreased consumption of growth factors. Results: HLCs demonstrated physiological function, e.g. albumin secretion, glycogen storage, urea production, indocyanin green and LDL uptake, collagen synthesis, and CYP activity. These cells were characterized and enriched based on LDL uptake as a biological property and infused intrasplencally to CCL4 treated mice model. Transplanted cells engrafted successfully and increased the survival rate. These cells remained functional after homing and promoted regeneration mechanisms in HLC transplanted group. Conclusion: This practical approach in scaling up and purifying strategy may help scientists to introduce promising methods for cell-based therapeutics and other biomedical application of HLCs.
- Subjects
PEPTIDES; C-terminal residues; STEM cell research; MEDICINE; CELL differentiation; PHYSIOLOGY
- Publication
Cell Journal (Yakhteh), 2013, Vol 15, Issue Sup 1, p28
- ISSN
2228-5806
- Publication type
Article