Physiological Microenvironmental Conditions in Different Scalable Culture Systems for Pluripotent Stem Cell Expansion and Differentiation

Abstract

Human Pluripotent Stem Cells (PSCs) are a valuable cell type that has a wide range of biomedical applications because they can differentiate into many types of adult somatic cell. Numerous studies have examined the clinical applications of PSCs. However, several factors such as bioreactor design, mechanical stress, and the physiological environment have not been optimized. These factors can significantly alter the pluripotency and proliferation properties of the cells, which are important for the mass production of PSCs. Nutritional mass transfer and oxygen transfer must be effectively maintained to obtain a high yield. Various culture systems are currently available for optimum cell propagation by maintaining the physiological conditions necessary for cell cultivation. Each type of culture system using a different configuration with various advantages and disadvantages affecting the mechanical conditions in the bioreactor, such as shear stress. These factors make it difficult to preserve the cellular viability and pluripotency of PSCs. Additional limitations of the culture system for PSCs must also be identified and overcome to maintain the culture conditions and enable large-scale expansion and differentiation of PSCs. This review describes the different physiological conditions in the various culture systems and recent developments in culture technology for PSC expansion and differentiation.

Hydrodynamic shear stress’ impact on mammalian cell properties and its applications in 3D bioprinting

“…The cellular microenvironment is another crucial factor that influences cell activity [159,[176][177][178]. Cell morphology alters significantly when floating in the medium as compared to adherent state [179].…”

Strategies for Generating Human Pluripotent Stem Cell- Derived-Organoid Culture for Disease Modelling, Drug Screening, and Regenerative Therapy

“…These materials from animal-derived components such as Matrigel or animal-derived-serum also may limit the therapeutic application of hPSC-derived organoids. In actual therapeutic applications in humans, this animal-origin substance may potentially induce the antigenicity and transmit a pathogen into hPSCs cultures, such as viruses [52].…”

“…These types of suspension culture enable better control of spheroid size and uniformity in a large amount at once, which influences the dynamic of hiPSCs expression factors of pluripotency and their differentiated counterparts [61]. The spheroid size and uniformity can be controlled by adjusting the inoculation density and manipulating the medium dynamics, such as rotation/stirring speed, fluidic flow, or medium volume [52], [62].…”

Particle-tracking-based estimation of shear stress on cell aggregates in shaking vessel with different shaking methods/speeds

“…Suspension culture is widely used for various purposes, such as the production of antibiotics, foods, and cells. Recently, various attempts have been made to mass-produce pluripotent stem cells as cell sources for regenerative medicine and drug screening 1,2 . Generally, a suspension culture system includes a dynamic system to agitate the medium for supplying oxygen, realizing uniform conditions in the medium, keeping cells oating, etc.…”