Regeneration
of Pancreatic Cells Using Optimized Nanoparticles
and l‑Glutamic Acid–Gelatin Scaffolds with
Controlled Topography and Grafted Activin A/BMP4
posted on 2023-11-13, 08:43authored byYung-Chih Kuo, Sheng-Yuan Lin, Sourav De, Rajendiran Rajesh
Regeneration of insulin-producing cells (IPCs) from induced
pluripotent
stem cells (iPSCs) under controlled conditions has a lot of promise
to emulate the pancreatic mechanism in vivo as a foundation of cell-based
diabetic therapy. l-Glutamic acid–gelatin scaffolds
with orderly pore sizes of 160 and 200 μm were grafted with
activin A and bone morphogenic proteins 4 (BMP4) to differentiate
iPSCs into definitive endoderm (DE) cells, which were then guided
with fibroblast growth factor 7 (FGF7)-grafted retinoic acid (RA)-loaded
solid lipid nanoparticles (FR-SLNs) to harvest IPCs. Response surface
methodology was adopted to optimize the l-glutamic acid-to-gelatin
ratio of scaffolds and to optimize surfactant concentration and lipid
proportion in FR-SLNs. Experimental results of immunofluorescence,
flow cytometry, and western blots revealed that activin A (100 ng/mL)–BMP4
(50 ng/mL)–l-glutamic acid (5%)–gelatin (95%)
scaffolds provoked the largest number of SOX17-positive DE cells from
iPSCs. Treatment with FGF7 (50 ng/mL)–RA (600 ng/mL)–SLNs
elicited the highest number of PDX1-positive β-cells from differentiated
DE cells. To imitate the natural pancreas, the scaffolds with controlled
topography were appropriate for IPC production with sufficient insulin
secretion. Hence, the current scheme using FR-SLNs and activin A–BMP4–l-glutamic acid–gelatin scaffolds in the two-stage differentiation
of iPSCs can be promising for replacing impaired β-cells in
diabetic management.