Chapter 7
Generation of Functional Insulin-Producing Cells from
Mouse Embryonic Stem Cells Through Protein Transduction
of Transcription Factors
Taku Kaitsuka and Kazuhito Tomizawa
Abstract
In this chapter, we describe a simple and unique method for the differentiation of mouse embryonic stem
cells into insulin-producing cells. In addition to cytokines and growth factors, key transcription factors for
pancreatic development are applied in this method through protein transduction technology. Furthermore,
a combination of nanofiber plates and laminin coatings improves the yield of differentiated cells. The
insulin-producing cells derived through this method express marker genes of mature β-cells and have an
ability to secrete insulin; therefore, these cells are useful for fundamental studies on pancreatic development,
drug development, and regenerative medicine for diabetes.
Key words Mouse embryonic stem cells, Pancreatic differentiation, Protein transduction, Insulinproducing cells, Insulin secretion
1 Introduction
Pluripotent stem cells, such as embryonic stem (ES) and induced
pluripotent stem (iPS) cells, are capable of differentiating into
somatic cells in vitro. Among these cells, pancreatic β-cells differentiated from pluripotent stem cells are useful for studies on pancreatic development, drug development, and regenerative medicine
for diabetes. To date, various protocols for pancreatic differentiation have been reported and utilized for basic and applied research
[1–4]. However, some of these protocols consist of numerous
complicated steps and require many hormones, growth factors,
and cytokines to guide the differentiation of the cells into insulinproducing cells.
We have established a virus-free method for pancreatic differentiation from mouse embryonic stem cells that is also simpler than
the previous protocols [5–7]. In stage 1 of our method, the cells are
treated with Activin A and basic fibroblast growth factor (bFGF)
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_7, © Springer Science+Business Media, LLC, part of Springer Nature 2021
85
Generation of Functional Insulin-Producing Cells from
Mouse Embryonic Stem Cells Through Protein Transduction
of Transcription Factors
Taku Kaitsuka and Kazuhito Tomizawa
Abstract
In this chapter, we describe a simple and unique method for the differentiation of mouse embryonic stem
cells into insulin-producing cells. In addition to cytokines and growth factors, key transcription factors for
pancreatic development are applied in this method through protein transduction technology. Furthermore,
a combination of nanofiber plates and laminin coatings improves the yield of differentiated cells. The
insulin-producing cells derived through this method express marker genes of mature β-cells and have an
ability to secrete insulin; therefore, these cells are useful for fundamental studies on pancreatic development,
drug development, and regenerative medicine for diabetes.
Key words Mouse embryonic stem cells, Pancreatic differentiation, Protein transduction, Insulinproducing cells, Insulin secretion
1 Introduction
Pluripotent stem cells, such as embryonic stem (ES) and induced
pluripotent stem (iPS) cells, are capable of differentiating into
somatic cells in vitro. Among these cells, pancreatic β-cells differentiated from pluripotent stem cells are useful for studies on pancreatic development, drug development, and regenerative medicine
for diabetes. To date, various protocols for pancreatic differentiation have been reported and utilized for basic and applied research
[1–4]. However, some of these protocols consist of numerous
complicated steps and require many hormones, growth factors,
and cytokines to guide the differentiation of the cells into insulinproducing cells.
We have established a virus-free method for pancreatic differentiation from mouse embryonic stem cells that is also simpler than
the previous protocols [5–7]. In stage 1 of our method, the cells are
treated with Activin A and basic fibroblast growth factor (bFGF)
Kumaran Narayanan (ed.), Bio-Carrier Vectors: Methods and Protocols, Methods in Molecular Biology, vol. 2211,
https://doi.org/10.1007/978-1-0716-0943-9_7, © Springer Science+Business Media, LLC, part of Springer Nature 2021
85