Enhanced differentiation of human amniotic fluid‐derived stem cells into insulin‐producing cells in vitro

Abstract

Aims/IntroductionTo investigate the ability of human amniotic fluid stem cells (hAFSCs) to differentiate into insulin‐producing cells.Materials and Methods hAFSCs were induced to differentiate into pancreatic cells by a multistep protocol. The expressions of pancreas‐related genes and proteins, including pancreatic and duodenal homeobox‐1, insulin, and glucose transporter 2, were detected by polymerase chain reaction and immunofluorescence. Insulin secreted from differentiated cells was tested by enzyme‐linked immunosorbent assay.Results hAFSCs were successfully isolated from amniotic fluid that expressed the pluripotent markers of embryonic stem cells, such as Oct3/4, and mesenchymal stem cells, such as integrin β‐1 and ecto‐5′‐nucleotidase. Here, we first obtained the hAFSCs that expressed pluripotent marker stage‐specific embryonic antigen 1. Real‐time polymerase chain reaction analysis showed that pancreatic and duodenal homeobox‐1, paired box gene 4 and paired box gene 6 were expressed in the early phase of induction, and then stably expressed in the differentiated cells. The pancreas‐related genes, such as insulin, glucokinase, glucose transporter 2 and Nkx6.1, were expressed in the differentiated cells. Immunofluorescence showed that these differentiated cells co‐expressed insulin, C‐peptide, and pancreatic and duodenal homeobox‐1. Insulin was released in response to glucose stimulation in a manner similar to that of adult human islets.ConclusionsThe present study showed that hAFSCs, under selective culture conditions, could differentiate into islet‐like insulin‐producing cells, which might be used as a potential source for transplantation in patients with type 1 diabetes mellitus.

Current progress in stem cell therapy for type 1 diabetes mellitus

“…In contrast, iPSCs are derived from adult somatic cells that have been reprogrammed back into an embryonic-like pluripotent state using Yamanaka factors [16,17]. During the last two decades, numerous methods to generate IPCs from hPSCs have been reported [9][10][11][12][18][19][20][21][22].…”

“…Current strategies for generating IPCs are mainly based on approaches that mimic normal pancreas development. The obtained IPCs are supposed to express specific biological markers of normal β cells that identify a terminal differentiation status, such as MAFA (a basic leucine zipper transcription factor expressed in mature β cells and absent in pancreatic progenitors and other cell types), NEUROD1 (downstream factor of NGN3 expressed in most pancreatic endocrine cells, including β cells), and PDX1/NKX 6.1 (restricted coexpression in β cells), as well as key functional features of adult β cells, including glucose-stimulated insulin secretion (GSIS) and C-peptide secretion [9][10][11][12][13][14]. In addition, after implantation into DM patients or immunodeficient diabetic animals, these in vitro-generated IPCs or islet organoids should respond to changing blood glucose and produce sufficient insulin and finally reverse hyperglycemia.…”

The combined effect of Pdx1 overexpression and Shh manipulation on the function of insulin‐producing cells derived from adipose‐tissue stem cells

“…The differentiated cells obtained exhibited a remarkable increase in insulin synthesis and secretion. These results revealed the usefulness of Shh pathway manipulation in improving IPC maturity . This study was therefore conducted to examine the effect of Pdx1 overexpression concurrent with Shh manipulation on differentiation outcomes.…”

The therapeutic applications of mesenchymal stromal cells from human perinatal tissues in autoimmune diseases

“…The insulin-producing cells can release insulin in a glucose-responsive manner and normalize hyperglycemia for a long period without immunosuppressive agents [ 98 , 99 ]. In addition to AMSCs, human AFSCs can also be induced to differentiate into insulin-producing cells; the induced AFSCs can secret insulin in response to glucose stimulation just like intrinsic pancreatic β cells [ 100 ].…”