Expansion Culture of Human Pluripotent Stem Cells and Production of Cardiomyocytes

Abstract

Transplantation of human pluripotent stem cell (hPSCs)-derived cardiomyocytes for the treatment of heart failure is a promising therapy. In order to implement this therapy requiring numerous cardiomyocytes, substantial production of hPSCs followed by cardiac differentiation seems practical. Conventional methods of culturing hPSCs involve using a 2D culture monolayer that hinders the expansion of hPSCs, thereby limiting their productivity. Advanced culture of hPSCs in 3D aggregates in the suspension overcomes the limitations of 2D culture and attracts immense attention. Although the hPSC production needs to be suitable for subsequent cardiac differentiation, many studies have independently focused on either expansion of hPSCs or cardiac differentiation protocols. In this review, we summarize the recent approaches to expand hPSCs in combination with cardiomyocyte differentiation. A comparison of various suspension culture methods and future prospects for dynamic culture of hPSCs are discussed in this study. Understanding hPSC characteristics in different models of dynamic culture helps to produce numerous cells that are useful for further clinical applications.

Stem Cell Therapy of Myocardial Infarction: A Promising Opportunity in Bioengineering

“…A large‐scale hPSC expansion system is required for more efficient and scalable production of hPSC‐CMs. Various suspension cultures of hPSCs have been developed with microcarriers, hydrogels, and polymers as supporting matrices that increase the production capacity ( Table ) . Other approaches for forming hPSC spheroids involve self‐aggregation in the absence of supporting materials.…”

Selection of human induced pluripotent stem cells lines optimization of cardiomyocytes differentiation in an integrated suspension microcarrier bioreactor

“…The differentiation of hPSCs/hiPSCs aggregates with and without MCs or with hydrogel [29] to CMs was also well demonstrated [37]. The scale up of the cardiac differentiation process into a fully closed system has also been well documented [37], e.g. in wave bag reactors [12] and with stirring tank reactors [27] as suspended embryoid bodies (EBs).…”

“…The expansion of hPSCs/hiPSCs in a spinner flask as aggregates with and without microcarriers (MCs) has been well documented [1,6,35] and scaled up into closed bioreactor systems [30,54] which paved the way to commercialization and industrial manufacturing [50]. The differentiation of hPSCs/hiPSCs aggregates with and without MCs or with hydrogel [29] to CMs was also well demonstrated [37]. The scale up of the cardiac differentiation process into a fully closed system has also been well documented [37], e.g.…”

“…However, it is suggested that doses of up to 1 billion functional CMs are required for a single cell therapy for an infarcted myocardium [15,16,24]. It has been a challenge to produce large quantities of high-purity CMs due to the adherent characteristic of stem cells and their differentiation efficiency [2,10,22,37]. A prerequisite for realizing the therapeutic potential of stem cells is the translation of inefficient manually operated monolayer base cell expansion and cardiac differentiation methods into an efficient, scalable, streamlined, closed system and controllable manufacturing process.…”

“…in wave bag reactors [12] and with stirring tank reactors [27] as suspended embryoid bodies (EBs). However, the processes of making EBs are labour intensive and problematic since it requires extensive cell handling such as dissociating or cutting [31,37]. Besides, although EBs with controlled size can be form by many methods [45,46,62], they might be limited to research scale [37].…”