Optimizing cell viability in droplet-based cell deposition

Hendriks, Jan; Visser, Claas Willem; Henke, Sieger; Leijten, Jeroen; Saris, Daniël B.F.; Sun, Chao; Lohse, Detlef; Karperien, Marcel

doi:10.1038/srep11304

Cited by 99 publications

(94 citation statements)

References 49 publications

(76 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the literature is described that the 3D‐bioprinting process can rupture or damage the membranes of cells, which sinks the viability rates and the success of bioprinting . The parameters applied during inkjet printing (e.g., pressure, valve nozzle size) can influence not only cell viability but also cell remodeling and differentiation capabilities .…”

Section: Discussionmentioning

confidence: 99%

Bioprinting Organotypic Hydrogels with Improved Mesenchymal Stem Cell Remodeling and Mineralization Properties for Bone Tissue Engineering

Campos

Blaeser

Buellesbach

et al. 2016

Adv Healthcare Materials

150

View full text Add to dashboard Cite

3D-manufactured hydrogels with precise contours and biological adhesion motifs are interesting candidates in the regenerative medicine field for the culture and differentiation of human bone-marrow-derived mesenchymal stem cells (MSCs). 3D-bioprinting is a powerful technique to approach one step closer the native organization of cells. This study investigates the effect of the incorporation of collagen type I in 3D-bioprinted polysaccharide-based hydrogels to the modulation of cell morphology, osteogenic remodeling potential, and mineralization. By combining thermo-responsive agarose hydrogels with collagen type I, the mechanical stiffness and printing contours of printed constructs can be improved compared to pure collagen hydrogels which are typically used as standard materials for MSC osteogenic differentiation. The results presented here show that MSC not only survive the 3D-bioprinting process but also maintain the mesenchymal phenotype, as proved by live/dead staining and immunocytochemistry (vimentin positive, CD34 negative). Increased solids concentrations of collagen in the hydrogel blend induce changes in cell morphology, namely, by enhancing cell spreading, that ultimately contribute to enhanced and directed MSC osteogenic differentiation. 3D-bioprinted agarose-collagen hydrogels with high-collagen ratio are therefore feasible for MSC osteogenic differentiation, contrarily to low-collagen blends, as proved by two-photon microscopy, Alizarin Red staining, and real-time polymerase chain reaction.

show abstract

Section: Discussionmentioning

confidence: 99%

Bioprinting Organotypic Hydrogels with Improved Mesenchymal Stem Cell Remodeling and Mineralization Properties for Bone Tissue Engineering

Campos

Blaeser

Buellesbach

et al. 2016

Adv Healthcare Materials

150

View full text Add to dashboard Cite

show abstract

“…It enables the generation of complex, multicellular tissue structures (e.g., vascularized tissue constructs) that cannot be created otherwise. On the other hand, the dispensing process involves mechanical stress, thermal stress, or a combination of both and ultimately affects cell behavior …”

Section: Methodsmentioning

confidence: 99%

Controlling Shear Stress in 3D Bioprinting is a Key Factor to Balance Printing Resolution and Stem Cell Integrity

Blaeser

Campos

Puster

et al. 2015

Adv Healthcare Materials

630

568

View full text Add to dashboard Cite

in cell biology, [ 29,30 ] e.g., in cell signaling and protein expression. [31][32][33] For instance, it is reported that shear stress promotes maturation of megakaryocytes. [ 34 ] Moderate shear stress was found to have an infl uence on stem cell differentiation. [ 35 ] Excessive shear stress, in contrast, even dispatches cells by disrupting the membrane. These phenomena are even more crucial in bioprinting processes, where hydrogels of high viscosity and small nozzles are applied in an attempt to improve the fi nal printing resolution. Here, we show that both hydrogel viscosity and nozzle size directly affect shear stress. To prevent adverse cell response and printing-related cell death, it is essential to control the shear stress level, identify its most important drivers, and study the cell response upon different stress levels. We hypothesize that regulating shear stress and elucidating its impact would be of great use in balancing cell integrity and printing resolution.We present a microvalve-based bioprinting system for the manufacturing of high resolution, multi-material 3D structures ( Scheme 1 A). Applying a straightforward fl uid-dynamics model, we were able to precisely control shear stress at the nozzle site, which could be adjusted by varying the printing pressure, hydrogel viscosity, and the nozzle diameter. Using this system, we conducted a broad study on how cell viability and proliferation potential are affected by different levels of shear stress (Scheme 1 B). Generating complex, multi-material 3D-structures, we demonstrate that high-resolution printing at moderate, cell-friendly nozzle shear stress are not mutually exclusive.The printer used throughout this study comprised four microvalve-based print heads, each individually controllable and heatable, mounted to a three-axis robotic system ( Figure S1, Supporting Information). A metal stage that could be lowered into a container fi lled with a bi-phasic support liquid-perfl uorocarbon (PFC) and an aqueous crosslinker solution-was used as a printing platform that allowed for the manufacturing of macroscopic, multi-layered 3D structures ( Figure S1, Supporting Information). The presented printing system dispenses single drops of cell-hydrogel suspension by jetting using electromagnetic microvalves. Thus, cells are primarily exposed to mechanical stress in the form of shear stress. To describe the shear stress condition in the nozzle of the valve, we developed a fl uid dynamics model for transient fl ow of non-Newtonian fl uids (hydrogels) based on the Bernoulli equation for unsteady fl ow: Equation (

show abstract

“…overview of the typical per-nozzle throughput (i.e., flow rate) as a function of the droplet size (i.e., diameter) of the most used continuous droplet manufacturing techniques for cell-laden microgel fabrication is presented. The indicated production regimes are based on data points obtained from the following references: droplet microfluidics [60,[91][92][93][94][95][96][97][98], vibrating jet [43], jet cutting [99,100], inkjet [101][102][103], air-induced spraying [100,[104][105][106][107], and electrospraying [100,[108][109][110]. If possible, multiple distinct data points were obtained from the mentioned studies.…”

Section: Balancing Throughput and Sizementioning

confidence: 99%

Single-Cell Microgels: Technology, Challenges, and Applications

Kamperman

Karperien

Gac

et al. 2018

Trends in Biotechnology

Self Cite

View full text Add to dashboard Cite

Single-cell-laden microgels effectively act as the engineered counterpart of the smallest living building block of life: a cell within its pericellular matrix. Recent breakthroughs have enabled the encapsulation of single cells in sub-100-μm microgels to provide physiologically relevant microniches with minimal mass transport limitations and favorable pharmacokinetic properties. Single-cell-laden microgels offer additional unprecedented advantages, including facile manipulation, culture, and analysis of individual cell within 3D microenvironments. Therefore, single-cell microgel technology is expected to be instrumental in many life science applications, including pharmacological screenings, regenerative medicine, and fundamental biological research. In this review, we discuss the latest trends, technical challenges, and breakthroughs, and present our vision of the future of single-cell microgel technology and its applications.

show abstract

Optimizing cell viability in droplet-based cell deposition

Cited by 99 publications

References 49 publications

Bioprinting Organotypic Hydrogels with Improved Mesenchymal Stem Cell Remodeling and Mineralization Properties for Bone Tissue Engineering

Bioprinting Organotypic Hydrogels with Improved Mesenchymal Stem Cell Remodeling and Mineralization Properties for Bone Tissue Engineering

Controlling Shear Stress in 3D Bioprinting is a Key Factor to Balance Printing Resolution and Stem Cell Integrity

Single-Cell Microgels: Technology, Challenges, and Applications

Contact Info

Product

Resources

About