A Covalently Crosslinked Ink for Multimaterials Drop‐on‐Demand 3D Bioprinting of 3D Cell Cultures

2021 ◽  
pp. 2100125
Author(s):  
Robert H. Utama ◽  
Vincent T. G. Tan ◽  
Kristel C. Tjandra ◽  
Andrew Sexton ◽  
Duyen H. T. Nguyen ◽  
...  
Keyword(s):  
Cell Cultures ◽  
3D Bioprinting ◽  
On Demand ◽  
Drop On Demand ◽  
3D Cell Cultures

scholarly journals Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

2020 ◽  
Author(s):  
Robert H. Utama ◽  
Lakmali Atapattu ◽  
Aidan P. O’Mahony ◽  
Christopher M. Fife ◽  
Jongho Baek ◽  
...  
Keyword(s):  
Cell Cultures ◽  
High Throughput ◽  
Cell Number ◽  
Drug Dose ◽  
3D Bioprinting ◽  
Multicellular Spheroids ◽  
Drop On Demand ◽  
3D Cell Cultures ◽  
The Impact

Abstract3D in vitro cancer models are important therapeutic and biological discovery tools, yet formation of multicellular spheroids in a throughput and highly controlled manner to achieve robust and statistically relevant data, remains challenging. Here, we developed an enabling technology consisting of a bespoke drop-on-demand 3D bioprinter capable of high-throughput printing of 96-well plates of spheroids. 3D-multicellular spheroids are embedded inside a tissue-like matrix with precise control over size and cell number. Application of 3D bioprinting for high-throughput drug screening was demonstrated with doxorubicin. Measurements showed that IC50 values were sensitive to spheroid size, embedding and how spheroids conform to the embedding, revealing parameters shaping biological responses in these models. Our study demonstrates the potential of 3D bioprinting as a robust high-throughput platform to screen biological and therapeutic parameters.Significance StatementIn vitro 3D cell cultures serve as more realistic models, compared to 2D cell culture, for understanding diverse biology and for drug discovery. Preparing 3D cell cultures with defined parameters is challenging, with significant failure rates when embedding 3D multicellular spheroids into extracellular mimics. Here, we report a new 3D bioprinter we developed in conjunction with bioinks to allow 3D-multicellular spheroids to be produced in a high-throughput manner. High-throughput production of embedded multicellular spheroids allowed entire drug-dose responses to be performed in 96-well plate format with statistically relevant numbers of data points. We have deconvoluted important parameters in drug responses including the impact of spheroid size and embedding in an extracellular matrix mimic on IC50 values.

scholarly journals Deep learning guided image-based droplet sorting for on-demand selection and analysis of single cells and 3D cell cultures

Lab on a Chip ◽  
2020 ◽  
Vol 20 (5) ◽  
pp. 889-900 ◽  
Author(s):  
Vasileios Anagnostidis ◽  
Benjamin Sherlock ◽  
Jeremy Metz ◽  
Philip Mair ◽  
Florian Hollfelder ◽  
...  
Keyword(s):  
Neural Networks ◽  
Deep Learning ◽  
Cell Cultures ◽  
Mammalian Cells ◽  
Single Cells ◽  
Bright Field ◽  
On Demand ◽  
3D Cell Cultures ◽  
Droplet Sorting ◽  
Demand Selection

To uncover the heterogeneity of cellular populations and multicellular constructs we show on-demand isolation of single mammalian cells and 3D cell cultures by coupling bright-field microdroplet imaging with real-time classification and sorting using convolutional neural networks.

scholarly journals Rheological Investigation of Collagen, Fibrinogen, and Thrombin Solutions for Drop-on-Demand 3D Bioprinting

2020 ◽  
Author(s):  
Hemanth Gudapati ◽  
Daniele Parisi ◽  
Ralph H. Colby ◽  
Ibrahim Ozbolat
Keyword(s):  
Bulk Viscosity ◽  
Flow Behavior ◽  
Three Dimensional ◽  
Protein Solutions ◽  
Ionic Surfactant ◽  
3D Bioprinting ◽  
Rheological Measurements ◽  
On Demand ◽  
Drop On Demand ◽  
Air Interface

<p>Collagen, fibrinogen, and thrombin proteins in aqueous buffer solutions are widely used as precursors of natural biopolymers for three-dimensional (3D) bioprinting applications. The proteins are sourced from animals and their quality may vary from batch to batch, inducing differences in the rheological properties of such solutions. In this work, we investigate the rheological response of collagen, fibrinogen, and thrombin protein solutions in bulk and at the solution/air interface. Interfacial rheological measurements show that fibrous collagen, fibrinogen and globular thrombin proteins adsorb and aggregate at the solution/air interface, forming a viscoelastic solid film at the interface. The viscoelastic film corrupts the bulk rheological measurements in rotational rheometers by contributing to an apparent yield stress, which increases the apparent bulk viscosity up to shear rates as high as 1000 s<sup>-1</sup>. The addition of a non-ionic surfactant, such as polysorbate 80 (PS80) in small amounts between 0.001 and 0.1 v/v%, prevents the formation of the interfacial layer, allowing the estimation of true bulk viscosity and viscoelastic properties of the solutions. The estimation of viscosity not only helps in identifying those protein solutions that are potentially printable with drop-on-demand (DOD) inkjet printing but also detects inconsistencies in flow behavior among the batches.</p>

scholarly journals A Novel 3D Bioprinter Using Direct-Volumetric Drop-On-Demand Technology for Fabricating Micro-Tissues and Drug-Delivery

2020 ◽  
Vol 21 (10) ◽  
pp. 3482 ◽  
Author(s):  
Brian E. Grottkau ◽  
Zhixin Hui ◽  
Yonggang Pang
Keyword(s):  
Drug Delivery ◽  
Cell Viability ◽  
High Throughput ◽  
Bone Cancer ◽  
Cell Types ◽  
High Viscosity ◽  
3D Bioprinting ◽  
On Demand ◽  
Drop On Demand ◽  
Short Period

Drop-on-demand (DOD) 3D bioprinting technologies currently hold the greatest promise for generating functional tissues for clinical use and for drug development. However, existing DOD 3D bioprinting technologies have three main limitations: (1) droplet volume inconsistency; (2) the ability to print only bioinks with low cell concentrations and low viscosity; and (3) problems with cell viability when dispensed under high pressure. We report our success developing a novel direct-volumetric DOD (DVDOD) 3D bioprinting technology that overcomes each of these limitations. DVDOD can produce droplets of bioink from <10 nL in volume using a direct-volumetric mechanism with <± 5% volumetric percent accuracy in an accurate spatially controlled manner. DVDOD has the capability of dispensing bioinks with high concentrations of cells and/or high viscosity biomaterials in either low- or high-throughput modes. The cells are subjected to a low pressure during the bioprinting process for a very short period of time that does not negatively impact cell viability. We demonstrated the functions of the bioprinter in two distinct manners: (1) by using a high-throughput drug-delivery model; and (2) by bioprinting micro-tissues using a variety of different cell types, including functional micro-tissues of bone, cancer, and induced pluripotent stem cells. Our DVDOD technology demonstrates a promising platform for generating many types of tissues and drug-delivery models.

scholarly journals Rheological investigation of collagen, fibrinogen, and thrombin solutions for drop-on-demand 3D bioprinting

Soft Matter ◽  
2020 ◽  
Vol 16 (46) ◽  
pp. 10506-10517 ◽  
Author(s):  
Hemanth Gudapati ◽  
Daniele Parisi ◽  
Ralph H. Colby ◽  
Ibrahim T. Ozbolat
Keyword(s):  
Three Dimensional ◽  
3D Bioprinting ◽  
Aqueous Buffer ◽  
On Demand ◽  
Buffer Solutions ◽  
Drop On Demand

Collagen, fibrinogen, and thrombin proteins in aqueous buffer solutions are widely used as precursors of natural biopolymers in three-dimensional (3D) bioprinting applications.

Rheological Investigation of Collagen, Fibrinogen, and Thrombin Solutions for Drop-on-Demand 3D Bioprinting

2020 ◽  
Author(s):  
Hemanth Gudapati ◽  
Daniele Parisi ◽  
Ralph H. Colby ◽  
Ibrahim Ozbolat
Keyword(s):  
Bulk Viscosity ◽  
Flow Behavior ◽  
Three Dimensional ◽  
Protein Solutions ◽  
Ionic Surfactant ◽  
3D Bioprinting ◽  
Rheological Measurements ◽  
On Demand ◽  
Drop On Demand ◽  
Air Interface

<p>Collagen, fibrinogen, and thrombin proteins in aqueous buffer solutions are widely used as precursors of natural biopolymers for three-dimensional (3D) bioprinting applications. The proteins are sourced from animals and their quality may vary from batch to batch, inducing differences in the rheological properties of such solutions. In this work, we investigate the rheological response of collagen, fibrinogen, and thrombin protein solutions in bulk and at the solution/air interface. Interfacial rheological measurements show that fibrous collagen, fibrinogen and globular thrombin proteins adsorb and aggregate at the solution/air interface, forming a viscoelastic solid film at the interface. The viscoelastic film corrupts the bulk rheological measurements in rotational rheometers by contributing to an apparent yield stress, which increases the apparent bulk viscosity up to shear rates as high as 1000 s<sup>-1</sup>. The addition of a non-ionic surfactant, such as polysorbate 80 (PS80) in small amounts between 0.001 and 0.1 v/v%, prevents the formation of the interfacial layer, allowing the estimation of true bulk viscosity and viscoelastic properties of the solutions. The estimation of viscosity not only helps in identifying those protein solutions that are potentially printable with drop-on-demand (DOD) inkjet printing but also detects inconsistencies in flow behavior among the batches.</p>

scholarly journals A covalently crosslinked bioink for multi-materials drop-on-demand 3D bioprinting of three-dimensional cell cultures

2021 ◽  
Author(s):  
Robert H. Utama ◽  
Vincent T. G. Tan ◽  
Kristel C. Tjandra ◽  
Andrew Sexton ◽  
Duyen H. T. Nguyen ◽  
...  
Keyword(s):  
Cell Culture ◽  
Human Error ◽  
Three Dimensional ◽  
Polymer System ◽  
Ductal Adenocarcinoma ◽  
3D Bioprinting ◽  
Cell Models ◽  
On Demand ◽  
Drop On Demand

AbstractIn vitro three-dimensional (3D) cell models have been accepted to better recapitulate aspects of in vivo organ environment than 2D cell culture. Currently, the production of these complex in vitro 3D cell models with multiple cell types and microenvironments remains challenging and prone to human error. Here we report a versatile bioink comprised of a 4-arm PEG based polymer with distal maleimide derivatives as the main ink component and a bis-thiol species as the activator that crosslinks the polymer to form the hydrogel in less than a second. The rapid gelation makes the polymer system compatible with 3D bioprinting. The ink is combined with a drop-on-demand 3D bioprinting platform consisting of eight independently addressable nozzles and high-throughput printing logic for creating complex 3D cell culture models. The combination of multiple nozzles and fast printing logic enables the rapid preparation of many complex 3D structures comprising multiple hydrogel environments in the one structure in a standard 96-well plate format. The platform compatibility for biological applications was validated using pancreatic ductal adenocarcinoma cancer (PDAC) cells with their phenotypic responses controlled by tuning the hydrogel microenvironment.

scholarly journals Can keratin scaffolds be used for creating three-dimensional cell cultures?

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 249-253
Author(s):  
Marta Bochynska-Czyz ◽  
Patrycja Redkiewicz ◽  
Hanna Kozlowska ◽  
Joanna Matalinska ◽  
Marek Konop ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Chemical Activation ◽  
Three Dimensional ◽  
Enzymatic Digestion ◽  
Pepsin Digestion ◽  
Cell Culturing ◽  
3D Cell Cultures ◽  
Associated Proteins ◽  
Positive Effect ◽  
Dimensional Cell

AbstractThree-dimensional (3D) cell cultures were created with the use of fur keratin associated proteins (F-KAPs) as scaffolds. The procedure of preparation F-KAP involves combinations of chemical activation and enzymatic digestion. The best result in porosity and heterogeneity of F-KAP surface was received during pepsin digestion. The F-KAP had a stable structure, no changes were observed after heat treatment, shaking and washing. The 0.15-0.5 mm fraction had positive effect for formation of 3D scaffolds and cell culturing. Living rat mesenchymal cells on the F-KAP with no abnormal morphology were observed by SEM during 32 days of cell culturing.

scholarly journals Correction: Assessment of the toxicity and inflammatory effects of different-sized zinc oxide nanoparticles in 2D and 3D cell cultures

RSC Advances ◽  
2020 ◽  
Vol 10 (72) ◽  
pp. 44397-44397
Author(s):  
Zhipan Wu ◽  
Rongfa Guan ◽  
Miao Tao ◽  
Fei Lyu ◽  
Guozhou Cao ◽  
...  
Keyword(s):  
Zinc Oxide ◽  
Cell Cultures ◽  
Zinc Oxide Nanoparticles ◽  
Oxide Nanoparticles ◽  
3D Cell Cultures ◽  
2D And 3D

Correction for ‘Assessment of the toxicity and inflammatory effects of different-sized zinc oxide nanoparticles in 2D and 3D cell cultures’ by Zhipan Wu, Rongfa Guan, Miao Tao et al., RSC Adv., 2017, 7, 12437–12445, DOI: 10.1039/C6RA27334C.

Sign in / Sign up

Export Citation Format

Share Document