Embryonic stem cells (ESC) are totipotent cells that can differentiate into a large number of different cell types. Stem cell‐derived, differentiated cells are of increasing importance as a potential source for non‐proliferating cells (e.g., cardiomyocytes or neurons) for future tissue engineering applications. Differentiation of ESC is initiated by the formation of embryoid bodies (EB). Current protocols for the generation of EB are either of limited productivity or deliver EB with a large variation in size and differentiation state. To establish an efficient and robust EB production process, we encapsulated mouse ESC into alginate microbeads using various microencapsulation technologies. Microencapsulation and culturing of ESC in 1.1% alginate microbeads gives rise to discoid colonies, which further differentiate within the beads to cystic EB and later to EB containing spontaneously beating areas. However, if ESC are encapsulated into 1.6% alginate microbeads, differentiation is inhibited at the morula‐like stage, so that no cystic EB can be formed within the beads. ESC colonies, which are released from 1.6% alginate microbeads, can further differentiate to cystic EB with beating cardiomyocytes. Extended supplementation of the growth medium with retinoic acid promotes differentiation to smooth muscle cells.
Engineering of functional tissues is a fascinating and fertile arena of research and development. This flourishing enterprise weaves together many areas of research to tackle the most complex question faced to date, namely how to design and reconstruct a synthetic three‐dimensional fully functional tissue on demand. At present our healthcare is under threat by several social and economical issues together with those of a more scientific and clinical nature. One such issue arises from our increasing life expectancy, resulting in an ageing society. This steeply growing ageing society requires functional organotypic tissues on demand for repair, replacement, and rejuvenation (R3). Several approaches are pioneered and developed to assist conventional tissue/organ transplantation. In this Progress Report, “non‐contact jet‐based” approaches for engineering functional tissues are introduced and bio‐electrosprays and cell electrospinning, i.e., biotechniques that have demonstrated as being benign for directly handling living cells and whole organisms, are highlighted. These biotechniques possess the ability to directly handle heterogeneous cell populations as suspensions with a biopolymer and/or other micro/nanomaterials for directly forming three‐dimensional functional living reconstructs. These discoveries and developments have provided a promising biotechnology platform with far‐reaching ramifications for a wide range of applications in basic biological laboratories to their utility in the clinic.
Electrospinning, a flexible jet-based fiber, scaffold, and membrane fabrication approach, has been elucidated as having significance to the heath sciences. Its capabilities have been most impressive as it possesses the ability to spin composite fibers ranging from the nanometer to the micrometer scale. Nonetheless, electrospinning has limitations and hazards, negating its wider exploration, for example, the inability to handle highly conducting suspensions, to its hazardous high voltage. Hence, to date electrospinning has undergone an exhaustive research regime to a point of cliché. Thus, in the work reported herein we unveil a competing technique to electrospinning, which has overcome the above limitations and hazards yet comparable in capabilities. The fiber preparation approach unearthed herein is referred to as "pressure driven spinning ͑PDS͒." The driving mechanism exploited in this fiber spinning process is the pressurized by-pass flow. This mechanism allows the drawing of either micro-or nanosized fibers while processing polymeric suspensions containing a wide range of advanced materials spanning structural, functional, and biological entities. Similar to electrospinning if the collection time of these continuous formed fibers is varied, composite scaffolds and membranes are generated. In keeping with our interests, multicompositional structural entities such as these could have several applications in biology and medicine, for example, ranging from the development of three-dimensional cultures ͑including disease models͒ to the development of synthetic tissues and organ structures to advanced approaches for controlled and targeted therapeutics.
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