Abstract:Tissue engineering requires the precise positioning of mammalian cells and biomaterials on substrate surfaces or in preprocessed scaffolds. Although the development of 2D and 3D bioprinting technologies has made substantial progress in recent years, precise, cell‐friendly, easy to use, and fast technologies for selecting and positioning mammalian cells with single cell precision are still in need. A new laser‐based bioprinting approach is therefore presented, which allows the selection of individual cells from… Show more
“…Under appropriate conditions, single cells can be reliably transferred with a high survival rate after transfer through the entire pulse duration range. This agrees very well with our previous findings obtained for a laser pulse duration of 0.6 ps [22,23]. Nevertheless, it is mandatory to investigate the long-term cell viability of ps-laser printed cells in a future study.…”
Section: Introductionsupporting
confidence: 92%
“…Due to the lower mass-density, living human mesenchymal stem cells (hMSC, SCP1 cell line) were suspended at the hydrogel surface (Figure 1 left). To visualize and investigate the process dynamics, including cavitation bubble expansion and subsequent jet propagation, pump-probe [30] time-resolved imaging observation was carried out, as described previously [23]. Briefly, the transfer process was illuminated by a 28 ns white-light pulse (Nanolite KL-L, High-Speed Photo-Systeme, Wedel, Germany) and then was captured with an imaging system comprising a microscope objective (Mitutoyo Plan Apo 5×/0.14, Japan), a tube lens (TTL200-A, Thorlabs, Bergkirchen, Germany), and a CCD1 camera (PCO, Pixefly USB, Kelheim, Germany).…”
Section: Methodsmentioning
confidence: 99%
“…With increasing laser pulse duration, we observed less efficient energy conversion and comparable, but slower jet dynamics. In our earlier work the cell survival rate obtained immediately after the transfer showed no significant difference in their ability to migrate and proliferate over a 66 h time scale [22,23]. Thus, to test the principal bioprinting compatibility at different pulse durations, only the cell survival rate 15 min after transfer was determined.…”
Section: Introductionmentioning
confidence: 99%
“…In our previous work [22,23], we demonstrated that by focusing a 600 fs laser pulse with a wavelength of 1030 nm into a hydrogel underneath a layer of cells, small droplets of hydrogel containing living cells can be ejected from the reservoir and transferred to an acceptor stage. The transferred cells showed a survival rate up to 95% without DNA double strand breaks and maintained their ability to migrate and proliferate even 66 h after the cell transfer.…”
Section: Introductionmentioning
confidence: 99%
“…Furthermore, the single cells can be transferred with a position precision of ±11.8 µm. By integrating this film-free cell transfer approach into an inverted epifluorescence microscope, this new approach allowed the precise and efficient selection and positioning of individual mammalian cells from a complex cell mixture (based on morphology or fluorescence) onto a 2D target substrate or a 3D pre-processed scaffold with single cell spatial precision [23]. It was found that the transfer process requires less than 100 µs per cell, manually searching for a cell takes ≈5 s, setting the laser focus another ≈5 s, and positioning of the acceptor surface takes further ≈10 s. To date the technique can be used for single cell studies.…”
Femtosecond laser pulses have been successfully used for film-free single-cell bioprinting, enabling precise and efficient selection and positioning of individual mammalian cells from a complex cell mixture (based on morphology or fluorescence) onto a 2D target substrate or a 3D pre-processed scaffold. In order to evaluate the effects of higher pulse durations on the bioprinting process, we investigated cavitation bubble and jet dynamics in the femto- and picosecond regime. By increasing the laser pulse duration from 600 fs to 14.1 ps, less energy is deposited in the hydrogel for the cavitation bubble expansion, resulting in less kinetic energy for the jet propagation with a slower jet velocity. Under appropriate conditions, single cells can be reliably transferred with a cell survival rate after transfer above 95% through the entire pulse duration range. More cost efficient and compact laser sources with pulse durations in the picosecond range could be used for film-free bioprinting and single-cell transfer.
“…Under appropriate conditions, single cells can be reliably transferred with a high survival rate after transfer through the entire pulse duration range. This agrees very well with our previous findings obtained for a laser pulse duration of 0.6 ps [22,23]. Nevertheless, it is mandatory to investigate the long-term cell viability of ps-laser printed cells in a future study.…”
Section: Introductionsupporting
confidence: 92%
“…Due to the lower mass-density, living human mesenchymal stem cells (hMSC, SCP1 cell line) were suspended at the hydrogel surface (Figure 1 left). To visualize and investigate the process dynamics, including cavitation bubble expansion and subsequent jet propagation, pump-probe [30] time-resolved imaging observation was carried out, as described previously [23]. Briefly, the transfer process was illuminated by a 28 ns white-light pulse (Nanolite KL-L, High-Speed Photo-Systeme, Wedel, Germany) and then was captured with an imaging system comprising a microscope objective (Mitutoyo Plan Apo 5×/0.14, Japan), a tube lens (TTL200-A, Thorlabs, Bergkirchen, Germany), and a CCD1 camera (PCO, Pixefly USB, Kelheim, Germany).…”
Section: Methodsmentioning
confidence: 99%
“…With increasing laser pulse duration, we observed less efficient energy conversion and comparable, but slower jet dynamics. In our earlier work the cell survival rate obtained immediately after the transfer showed no significant difference in their ability to migrate and proliferate over a 66 h time scale [22,23]. Thus, to test the principal bioprinting compatibility at different pulse durations, only the cell survival rate 15 min after transfer was determined.…”
Section: Introductionmentioning
confidence: 99%
“…In our previous work [22,23], we demonstrated that by focusing a 600 fs laser pulse with a wavelength of 1030 nm into a hydrogel underneath a layer of cells, small droplets of hydrogel containing living cells can be ejected from the reservoir and transferred to an acceptor stage. The transferred cells showed a survival rate up to 95% without DNA double strand breaks and maintained their ability to migrate and proliferate even 66 h after the cell transfer.…”
Section: Introductionmentioning
confidence: 99%
“…Furthermore, the single cells can be transferred with a position precision of ±11.8 µm. By integrating this film-free cell transfer approach into an inverted epifluorescence microscope, this new approach allowed the precise and efficient selection and positioning of individual mammalian cells from a complex cell mixture (based on morphology or fluorescence) onto a 2D target substrate or a 3D pre-processed scaffold with single cell spatial precision [23]. It was found that the transfer process requires less than 100 µs per cell, manually searching for a cell takes ≈5 s, setting the laser focus another ≈5 s, and positioning of the acceptor surface takes further ≈10 s. To date the technique can be used for single cell studies.…”
Femtosecond laser pulses have been successfully used for film-free single-cell bioprinting, enabling precise and efficient selection and positioning of individual mammalian cells from a complex cell mixture (based on morphology or fluorescence) onto a 2D target substrate or a 3D pre-processed scaffold. In order to evaluate the effects of higher pulse durations on the bioprinting process, we investigated cavitation bubble and jet dynamics in the femto- and picosecond regime. By increasing the laser pulse duration from 600 fs to 14.1 ps, less energy is deposited in the hydrogel for the cavitation bubble expansion, resulting in less kinetic energy for the jet propagation with a slower jet velocity. Under appropriate conditions, single cells can be reliably transferred with a cell survival rate after transfer above 95% through the entire pulse duration range. More cost efficient and compact laser sources with pulse durations in the picosecond range could be used for film-free bioprinting and single-cell transfer.
Artificial biological structures hold the promise for modeling cellular assembly in vitro and have advanced considerable studies in cell biology, disease modeling, drug testing, and regenerative medicine. Biological functions are derived from micro‐ and macroscale interactions of various cell types, and a structural property matching the tissue in vivo is required to enable precision biological function. Despite various types of tissues and organs are successfully constructed by conventional biofabrication technologies, they mostly only show a small fraction of structural features found in real tissues. Tools for single‐cell manipulation provide the approach to fabricate artificial tissues cell‐by‐cell, and have enabled the construction of biological structures with single‐cell and heterogeneous features, recapitulating the complexity in vivo. This review presents a comprehensive overview of the construction of biological structures through manipulating single cells, covering single‐cell technologies with operation principles and main advances, biological structures associated with informative explanations of single‐cell manipulation during construction, and representative applications mainly focusing on analysis and modeling. Current challenges and future perspectives in this field are also discussed.
Advancing biofabrication towards manufacturing living constructs with well‐defined architectures and increasingly biologically relevant cell densities is highly desired to mimic the biofunctionality of native human tissues. The formulation of tissue‐like, cell‐dense inks for biofabrication remains, however, challenging at various levels of the bioprinting process. Promising advances have been made towards this goal, and relatively high cell densities have been achieved, surpassing the limited cell densities of conventional platforms, pushing the current boundaries a step closer to achieving tissue‐like cell densities. On this focus, herein we discuss the overarching challenges in the bioprocessing of cell‐rich living inks into clinically‐grade engineered tissues, as well as highlight the most recent advances in cell‐rich living ink formulations and their processing technologies. Additionally, an overview of the foreseen developments in the field is provided and critically discussed.This article is protected by copyright. All rights reserved
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.