Insulin-Producing Cells Derived from Human Embryonic Stem Cells: Comparison of Definitive Endoderm- and Nestin-Positive Progenitor-Based Differentiation Strategies

Abstract

Human embryonic stem cells (hESCs) are pluripotent and capable of undergoing multilineage differentiation into highly specialized cells including pancreatic islet cells. Thus, they represent a novel alternative source for targeted therapies and regenerative medicine for diabetes. Significant progress has been made in differentiating hESCs toward pancreatic lineages. One approach is based on the similarities of pancreatic β cell and neuroepithelial development. Nestin-positive cells are selected as pancreatic β cell precursors and further differentiated to secrete insulin. The other approach is based on our knowledge of developmental biology in which the differentiation protocol sequentially reproduces the individual steps that are known in normal β cell ontogenesis during fetal pancreatic development. In the present study, the hESC cell line PKU1.1 was induced to differentiate into insulin-producing cells (IPCs) using both protocols. The differentiation process was dynamically investigated and the similarities and differences between both strategies were explored. Our results show that IPCs can be successfully induced with both differentiation strategies. The resulting IPCs from both protocols shared many similar features with pancreatic islet cells, but not mature, functional β cells. However, these differently-derived IPC cell types displayed specific morphologies and different expression levels of pancreatic islet development-related markers. These data not only broaden our outlook on hESC differentiation into IPCs, but also extend the full potential of these processes for regenerative medicine in diabetes.

Laminin 411 acts as a potent inducer of umbilical cord mesenchymal stem cell differentiation into insulin-producing cells

“…It was further confirmed in an Embryonic Stem Cells (ESCs) differentiation protocol [20]. So we want to know whether laminin 411 was through the same pathway to induce IPC generation.…”

Engineering Strategies for the Formation of Embryoid Bodies from Human Pluripotent Stem Cells

“…EBs formed in suspension cultures have generally demonstrated a high degree of heterogeneity in their downstream differentiation patterns [18], reducing the overall efficiency for directed differentiation into a specific lineage in high fidelity. Optimization of the differentiation protocols of EBs toward specific lineages requires the incorporation of the appropriate microengineering technology to obtain a monodisperse synchronous EB population that has a differentiation status enriched for desired lineages.…”

“…A wide distribution in the EB size introduces a source of variability in their downstream differentiation [17], which depends on the immediate microenvironment perceived by individual cells in the EBs, that is, the position of cells relative to others in the EBs. This effect is more pronounced when EBs exceed a certain size range: cells at the peripheral of the differentiating EBs tend to differentiate into the primitive endoderm, while the cells at the center of the EBs tend to give rise to primitive ectoderm cells [18]. When cultured in chondrogenic medium, small EBs exhibited higher propensity toward chondrogenesis, yet medium and large EBs shifted their potential toward hematopoietic and endothelial differentiation [19][20][21].…”

The Making of Pancreatic β Cells: Advances and Apprehensions

“…Rui Wei et al [122] compared the two routes of production, the first involving nestinpositive progenitors and second involving the definitive endoderm (DE), for the successful differentiation of cells into insulin producing cells (IPCs) from human embryonic stem cells (hESCs). In spite of the similarities of the results such as islet-like cell aggregation, expression of transcription factors such as Pdx1, MafA and Nkx6.1, production of pancreatic hormones such as Insulin, C-peptide and PP, Glucose -stimulated insulin production and IPC morphology among the both methods, there were certain differences observed (Table 4).…”

“…In addition to the report by Rui Wei et al [122], Mahmoud M.Gabr et al [123] compared three IPS protocols involved in the human bone marrow derived mesenchymal stem cells (HBM-MSC) production. The first protocol was the one-step protocol.…”