Developing a Cost-Effective and Scalable Production of Human Hepatic Competent Endoderm from Size-Controlled Pluripotent StemCell Aggregates

Publish Year: 1397
نوع سند: مقاله کنفرانسی
زبان: English
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NSCMRMED03_114

تاریخ نمایه سازی: 30 دی 1397

Abstract:

Background and Aim: Dynamic suspension culture of human pluripotentstem cells (hPSCs) in stirred bioreactors provides a valuable scalableculture platform for integrated differentiation toward different lineagesfor potential research and therapeutic applications. However, currentprotocols for scalable and integrated differentiation of hPSCs limited dueto the high cost of growth factors and technical challenges.Methods: hPSCs aggregates primed with 6 and 12 μM of CHIR99021(CHIR), a Wnt agonist, in combination with different concentrations ofhigh-cost Activin A (10, 25, 50, 100 ng/mL). We sought to determinethe appropriate treatment duration for efficient and cost-effectivedifferentiation protocol for foregut definitive endoderm production in adynamic suspension culture. Afterward, we evaluated the impact of theinitial hPSC aggregate sizes (small: 86±18 μm; medium: 142±32 μm;large: 214±34 μm) as critical bioprocess parameter on differentiationefficacy at the beginning of inductionResults: One-day priming of hPSCs as 3D aggregates (hPSpheres) with6 μM CHIR followed by treatment with a low concentration of Activin(10 ng/mL) for 2 days resulted in efficient differentiation to definitiveendoderm that highly expressed the anterior endodermal marker HEX.These endodermal cells differentiated efficiently into mature functionalhepatocytes. The medium-sized hPSpheres resulted in higher productivityand differentiation efficiency for scalable hepatocytes production,whereas small aggregates resulted in significant cell-loss after CHIRtreatment and large aggregates had less efficacious endodermaldifferentiation. Differentiated cells exhibited multiple characteristicsof primary hepatocytes as evidenced by expressions of liver-specificmarkers, indocyanine green (ICG) and low-density lipoprotein (LDL)uptake, and glycogen storageConclusion: This platform could be employed for the scalable productionof hPSC-derived hepatocytes for clinical and drug discovery applications.

Authors

Zahra Farzaneh

Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biologyand Technology, ACECR, Royan Institute, Tehran, Iran

Mostafa Najarasl

Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biologyand Technology, ACECR, Royan Institute, Tehran, Iran

Saeed Abbaszlizadeh

Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biologyand Technology, ACECR, Royan Institute, Tehran, Iran

Massoud Vosough

Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biologyand Technology, ACECR, Royan Institute, Tehran, Iran