Once you have your PSCs growing in culture and you have checked that they are expressing undifferentiated cell markers, you can start to differentiate your PSCs into your new cell type of interest. Due to the pluripotent nature of PSCs they have the potential to differentiate into any cell type from neural cells to cardiomyocytes. This differentiation can be done simply using a cocktail of different biochemicals and growth factors. Here we outline some of the biochemicals you can purchase on our catalog and the different cell types you can make using these.

Biochemicals

Neural cell types

SB431542, ALK inhibitor can be used in combination with Noggin to differentiate PSCs into neural progenitor cells (Chambers et al., 2009).

Cardiac cell types

5-Azacytidine has been shown to promote the differentiation of mesenchymal stem cells into cardiomyocytes (Makino et al., 1999).

Glycogen synthase kinase 3 (GSK-3) inhibitor, CHIR99021 can be used to differentiate PSCs into cardiomyocytes (Lian et al., 2013).

Hepatic cell types

Sodium butyrate can be used to generate hepatocytes from ESCs (Rambhatla et al., 2003).

Pancreatic cell types

RepSox, TGF-beta type 1 receptor inhibitor will induce PSCs to become pancreatic β-cells (Hosoya et al., 2012).

Growth factors and cytokines

Members of the fibroblast growth factor (FGF) family play key roles in the differentiation of stem cells into the 3 germ layers. Applying different FGF proteins in combination with other growth factors eg bone morphogenetic proteins (BMPs), Wnts, and epidermal growth factor (EGF) will lead to the differentiation of these germ layers into specific cell types. Here are some growth factor combinations for different cell types.

Neural cell types

FGF-2 and EGF can be combined to make neural progenitor cells from PSCs (Ostenfeld et al.,2004).

Cardiac cell types

BMP-2, BMP-4, Activin A, FGF, and Wnt5a all used in the differentiation of PSCs to cardiac progenitors (Rajala et al., 2011).

Pancreatic cell types

FGF-10 and FGF-7 are used to differentiate PCSs into pancreatic β cell progenitors (Shahjalal et al., 2018)

Hepatic cell types

Wnt3a, Activin A, FGF4, BMP-4, hepatocyte growth factor (HGF) are all used to generate liver progenitor cells (Du et al., 2018).

Lung cell types

BMP-4, Wnt3a, FGF-10, EGF and retinoic acid (RA) are all used in the production of alveolar type II-like epithelial cells from PSCs (Ghaedi et al., 2015).

References

1. Chambers, S. M.,, Fasano, C. A.,, et.al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling. Nature Biotechnology 27 (3),275-280 (2009)
2. Makino S, K , Fukuda, S, et. al. Cardiomyocytes can be generated from marrow stromal cells in vitro. J Clin Invest  103 ,697-705 (1999)
3. Lian, X.,, Zhang, J.,, etl. al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nature Protocols  8 (1),162-175 (2013)
4. Rambhatla, L.,, Chiu, C. P.,, Kundu, P.,, Peng, Y.,, & Carpenter, M. K. Generation of hepatocyte-like cells from human embryonic stem cells Cell Transplantation  12 (1),1-11 (2003)
5. Hosoya, M. Preparation of pancreatic β-cells from human iPS cells with small molecules. Islets  4 (3),249-252 (2012)
6. Ostenfeld T, , Svendsen CN. Requirement for neurogenesis to proceed through the division of neuronal progenitors following differentiation of epidermal growth factor and fibroblast growth factor-2-responsive human neural stem cells. Stem Cells  22 (5),798-811 (2000)
7. Shahjalal, H. M., , Abdal Dayem, , et. al. Generation of pancreatic β cells for treatment of diabetes: Advances and challenges. Stem Cell Research and Therapy,  9 (1), (2018)
8. Du, C., , Feng, Y., , Qiu, D., , et. al. Highly efficient and expedited hepatic differentiation from human pluripotent stem cells by pure small-molecule cocktails. Stem Cell Research and Therapy  9 (1),1-15 (2018)
9. Ghaedi, , M. Niklason L.E., , and Williams J Development of Lung Epithelium from Induced Pluripotent Stem Cells. Curr Transplant Rep.  2 (1),81-89 (2015)