Abstract:Understanding how microenvironmental cues influence cellular behavior will enable development of efficient and robust pluripotent stem cell differentiation protocols. Unlike traditional cell culture dishes, microfluidic bioreactors can provide stable microenvironmental conditions by continuous medium perfusion at a controlled rate. The aim of this study is to investigate whether a microfluidic culture device could be used as a perfused platform for long-term cell culture processes such as the retinal different… Show more
“…At the protein level, mESCs produced over 140 times more LIF inside microchambers than in standard 2D culture, where changes in microchamber height (volume) were used to regulate pluripotent phenotype of stem cells [272]. A more recent study produced the first long-term, continuously perfused microfluidic system for retinal differentiation of human induced pluripotent stem cells (hiPSCs) [273]. The perfusion flow rate was established via measurement of consumption/degradation of key growth factors significant to neural differentiation and survival, including insulin-like growth factor (IGF-1), and FGF.…”
Bioengineering systems have transformed scientific knowledge of cellular behaviors in the nervous system (NS) and pioneered innovative, regenerative therapies to treat adult neural disorders. Microscale systems with characteristic lengths of single to hundreds of microns have examined the development and specialized behaviors of numerous neuromuscular and neurosensory components of the NS. The visual system is comprised of the eye sensory organ and its connecting pathways to the visual cortex. Significant vision loss arises from dysfunction in the retina, the photosensitive tissue at the eye posterior that achieves phototransduction of light to form images in the brain. Retinal regenerative medicine has embraced microfluidic technologies to manipulate stem-like cells for transplantation therapies, where de/differentiated cells are introduced within adult tissue to replace dysfunctional or damaged neurons. Microfluidic systems coupled with stem cell biology and biomaterials have produced exciting advances to restore vision. The current article reviews contemporary microfluidic technologies and microfluidics-enhanced bioassays, developed to interrogate cellular responses to adult retinal cues. The focus is on applications of microfluidics and microscale assays within mammalian sensory retina, or neuro retina, comprised of five types of retinal neurons (photoreceptors, horizontal, bipolar, amacrine, retinal ganglion) and one neuroglia (Müller), but excludes the non-sensory, retinal pigmented epithelium.
“…At the protein level, mESCs produced over 140 times more LIF inside microchambers than in standard 2D culture, where changes in microchamber height (volume) were used to regulate pluripotent phenotype of stem cells [272]. A more recent study produced the first long-term, continuously perfused microfluidic system for retinal differentiation of human induced pluripotent stem cells (hiPSCs) [273]. The perfusion flow rate was established via measurement of consumption/degradation of key growth factors significant to neural differentiation and survival, including insulin-like growth factor (IGF-1), and FGF.…”
Bioengineering systems have transformed scientific knowledge of cellular behaviors in the nervous system (NS) and pioneered innovative, regenerative therapies to treat adult neural disorders. Microscale systems with characteristic lengths of single to hundreds of microns have examined the development and specialized behaviors of numerous neuromuscular and neurosensory components of the NS. The visual system is comprised of the eye sensory organ and its connecting pathways to the visual cortex. Significant vision loss arises from dysfunction in the retina, the photosensitive tissue at the eye posterior that achieves phototransduction of light to form images in the brain. Retinal regenerative medicine has embraced microfluidic technologies to manipulate stem-like cells for transplantation therapies, where de/differentiated cells are introduced within adult tissue to replace dysfunctional or damaged neurons. Microfluidic systems coupled with stem cell biology and biomaterials have produced exciting advances to restore vision. The current article reviews contemporary microfluidic technologies and microfluidics-enhanced bioassays, developed to interrogate cellular responses to adult retinal cues. The focus is on applications of microfluidics and microscale assays within mammalian sensory retina, or neuro retina, comprised of five types of retinal neurons (photoreceptors, horizontal, bipolar, amacrine, retinal ganglion) and one neuroglia (Müller), but excludes the non-sensory, retinal pigmented epithelium.
“…[11] Microfluidic performance exhibits many advantageous properties, such as rapid sample processing, spatiotemporal cell manipulation and analysis within precise and controlled microfluidic conditions. [12,13] Furthermore, microfluidic integration on a large-scale enables highthroughput operation and investigation of cell/tumor samples. [14,15] In the past several decades, research scholars around the globe have developed many microfluidic tumor manipulation systems that depend on passive microwells, [16][17][18][19] microdroplets, [20,21] microhydrogels, [22][23][24] active pneumatic microstructures (PμSs), [25][26][27] as well as 3D acoustic tweezers [28,29] for controllable 3D tumor cultivation.…”
Background: Cancer has been responsible for a large number of human deaths in the 21st century. Establishing a controllable, biomimetic, and large-scale analytical platform to investigate the tumor-associated pathophysiological and preclinical events, such as oncogenesis and chemotherapy, is necessary.
Methods and Results:This study presents antitumor investigation in a parallel, largescale, and tissue-mimicking manner based on well-constructed chemical gradients and heterotypic three-dimensional (3D) tumor cocultures using a multifunctionintegrated device. The integrated microfluidic device was engineered to produce a controllable and steady chemical gradient by manipulative optimization. Array-like and size-homogeneous production of heterotypic 3D tumor cocultures with in vivolike features, including similar tumor-stromal composition and functional phenotypic gradients of metabolic activity and viability, was successfully established. Furthermore, temporal, parallel, and high-throughput analyses of tumor behaviors in different antitumor stimulations were performed in a device based on the integrated operations involving gradient generation and coculture.
Conclusion:This achievement holds great potential for applications in the establishment of multifunctional tumor platforms to perform tissue-biomimetic neoplastic research and therapy assessment in the fields of oncology, bioengineering, and drug discovery.
“…Refreshing the culture medium removes these signaling factors. Consequently, the medium perfusion rate changes the overall mean level of cell-secreted factors, making it an important parameter in microfluidic stem cell culture (Titmarsh et al 2011 ; Yoshimitsu et al 2014 ; Abdolvand et al 2019 ; Fattahi et al 2020 ). The optimal perfusion rate, in turn, depends on the volume of the cell culture space.…”
Human stem cell-derived cells and tissues hold considerable potential for applications in regenerative medicine, disease modeling and drug discovery. The generation, culture and differentiation of stem cells in low-volume, automated and parallelized microfluidic chips hold great promise to accelerate the research in this domain. Here, we show that we can differentiate human embryonic stem cells (hESCs) to early cardiac mesodermal cells in microfluidic chambers that have a volume of only 30 nanoliters, using discontinuous medium perfusion. 64 of these chambers were parallelized on a chip which contained integrated valves to spatiotemporally isolate the chambers and automate cell culture medium exchanges. To confirm cell pluripotency, we tracked hESC proliferation and immunostained the cells for pluripotency markers SOX2 and OCT3/4. During differentiation, we investigated the effect of different medium perfusion frequencies on cell reorganization and the expression of the early cardiac mesoderm reporter MESP1mCherry by live-cell imaging. Our study demonstrates that microfluidic technology can be used to automatically culture, differentiate and study hESC in very low-volume culture chambers even without continuous medium perfusion. This result is an important step towards further automation and parallelization in stem cell technology.
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