Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells

Mobarra, N. and Soleimani, M. and Pakzad, R. and Enderami, S. E. and Pasalar, P. (2018) Three-dimensional nanofiberous PLLA/PCL scaffold improved biochemical and molecular markers hiPS cell-derived insulin-producing islet-like cells. Artificial Cells, Nanomedicine and Biotechnology, 46 (sup3). S685-S692.

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Abstract

Nanofibrous scaffolds are considered as a new strategy for Type 1 diabetes mellitus therapy. We used a hybrid of poly-l-lactic acid (PLLA) and polycaprolactone (PCL) as three-dimensional (3D) culture models for differentiation of human induced pluripotent stem cells (hiPSCs) to beta islet-like cluster cell compared with routine culture (2D). Morphological changes of cells were checked by microscope. mRNA endodermal SOX-17 on day 7 and pancreatic gene markers Pdx1, glucagon and Glut2 were evaluated on day 23 by qPCR. As well as, insulin and C-peptide protein expression was evaluated by immunocytochemistry staining. In addition, insulin and C-peptide secretion in various glucose concentrations was evaluated by ELISA. Light and scanning electron microscopy (SEM) microscope showed changes in induced cells. In tandem, these modifications were more evident in 3 D culture. Pdx1, Glucagon and Glut2 markers in PLLA/PCL were significantly higher in 3 D culture. In addition, qualitative immunochemistry showed that insulin and C-peptide were expressed in 2 D and 3 D culture medium. Furthermore, evaluation of insulin and C-peptide clarified that secretion of these proteins in PLLA/PCL scaffold were statistically different in 2 D and 3 D strategies. These findings suggest that functional matured induction cells on PLLA/PCL scaffold can be used for islet beta cell therapy and regenerative medicine. © 2018, © 2018 Informa UK Limited, trading as Taylor & Francis Group.

Item Type: Article
Additional Information: Cited By :5 Export Date: 16 February 2020 Correspondence Address: Soleimani, M.; Department of Hematology, School of Medical Sciences, Tarbiat Modares UniversityIran; email: soleimm@modares.ac.ir
Uncontrolled Keywords: C-peptide Human induced pluripotent stem cells Insulin PLLA/PCL scaffold Type 1 diabetes mellitus Biochemistry Cytology Lactic acid Peptides Physiology Polycaprolactone Polymer blends Polymerase chain reaction Scanning electron microscopy Stem cells Glucose concentration Human-induced pluripotent stem cells Immunocytochemistry Morphological changes Nanofibrous scaffolds PLLA Threedimensional (3-d) Scaffolds (biology) biochemical marker C peptide gelatin glucagon glucose glucose transporter 2 messenger RNA molecular marker molecular scaffold octamer transcription factor 4 polylactic acid transcription factor PDX 1 transcription factor Sox17 differentiation antigen nanofiber polyester polylactide Article cell culture technique cell differentiation cell structure controlled study embryoid body enzyme linked immunosorbent assay human human cell human cell culture hydrophilicity immunochemistry immunofluorescence test induced pluripotent stem cell insulin release marker gene pancreas islet beta cell pancreas secretion protein expression protein secretion quantitative analysis real time polymerase chain reaction stem cell culture three dimensional culture two dimensional culture biosynthesis chemistry metabolism tissue scaffold Antigens, Differentiation Humans Induced Pluripotent Stem Cells Insulin-Secreting Cells Nanofibers Polyesters Tissue Scaffolds
Subjects: WK Endocrine System
QU Biochemistry
Divisions: Mashhad University of Medical Sciences
Depositing User: lib2 lib2 lib2
Date Deposited: 08 May 2020 17:00
Last Modified: 08 May 2020 17:00
URI: http://eprints.mums.ac.ir/id/eprint/17402

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