Differentiation of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells to Cardiomyocytes

Abstract

Since human embryonic stem cells (hESCs) were first differentiated to beating cardiomyocytes a decade ago, interest in their potential applications has increased exponentially. This has been further enhanced over recent years by the discovery of methods to induce pluripotency in somatic cells, including those derived from patients with hereditary cardiac diseases. Human pluripotent stem cells have been among the most challenging cell types to grow stably in culture but advances in reagent development now mean that most laboratories can expand both embryonic and induced pluripotent stem cells robustly using commercially available products. However, differentiation protocols have lagged behind and, in many cases, only produce the cell types required with low efficiency. Cardiomyocyte differentiation techniques were also initially inefficient and not readily transferable across cell lines, but there are now a number of more robust protocols available. Here we review the basic biology underlying the differentiation of pluripotent cells to cardiac lineages and describe current state-of-the-art protocols as well as ongoing refinements. This should provide a useful entry for laboratories new to this area to start their research. Ultimately, efficient and reliable differentiation methodologies are essential to generate desired cardiac lineages in order to realize the full promise of human pluripotent stem cells for biomedical research, drug development, and clinical applications.

Engineering Cardiovascular Regeneration

“…They are based on Matrigel [66] or feeder-cell coated vessels. The differentiation efficiency is highly depending on the timing, differentiation factors, and culture medium (review in [71]). By now, efficient protocols result in up to 90 % pure cardiomyocytes without genetic selection (see above), demonstrating that differentiation is no longer a major bottleneck on the way to cardiac replacement therapy.…”

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

“…TeSR2 is commonly marketed as an improved version of TeSR1, but they are essentially the same. Some authors have reported preliminary results showing that hESC that have been maintained in mTeSR1 may not readily form embryoid bodies [98], but no adverse effects on cells grown in TeSR1 as monolayers have been noted so far. medium supplemented with B27.…”

“…Monolayer protocols for induction of differentiation along the cardiogenic lineage normally use RPMI/B27 medium with Matrigel or, when completely animal-free conditions are required, synthetic products such as the [68,98,102].…”

“…The cell surface marker stage-specific antigen-1 (SSEA1, LeX/SSEA1/CD15 ) is typically not expressed in hESC maintained in the undifferentiated state, but its expression is quickly upregulated in cells that have embarked on the differentiation route and may therefore be used as an early differentiation marker [98]. Two other surface markers that may be used to indicate early stages of mesodermal differentiation are N-CAM (neural cell adhesion molecule, CD56) and EpCAM (epithelial adhesion molecule, CD326) [98,174].…”

Microfluidic system for enhanced cardiac tissue formation

“…This was possible thanks to (1) generation of induced pluripotent stem cells (iPSCs) from somatic cells of the patient and (2) optimization of in vitro differentiation of iPSCs into cardiomyocytes (CMs) [1]. Currently, we are able to derive human chamber-specific CMs using robust and highly efficient protocols [2][3][4].…”