Scaffold-free bioprinting of mesenchymal stem cells with the regenova printer: Optimization of printing parameters

Aguilar, Izath Nizeet; Smith, Lester J.; Olivos, David J.; Chu, Tien Min G.; Kacena, Melissa A.; Wagner, Diane R.

doi:10.1016/j.bprint.2019.e00048

Cited by 38 publications

(42 citation statements)

References 9 publications

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“…The Regenova Bio 3D Printer (Cyfuse Biomedical K.K., Tokyo, Japan), a semi-automated biofabrication robot, was used in two distinct ways for the scope of this work. First, the Regenova Bio 3D Printer used a vision system to examine spheroids and assess roundness, surface smoothness, and diameter [29][30][31]. This assessment was used to optimize spheroid parameters before biofabrication because the Kenzan method requires a specific set of characteristics for successful biofabrication.…”

Section: Biofabrication Equipment and The Kenzan Methodsmentioning

confidence: 99%

“…In the Japanese form of flower arranging, a kenzan is a metal needle array used to hold floral arrangements in place ( Figure 1A). In the Kenzan method of biofabrication, the Kenzan is the microneedle array ( Figure 1B) used to hold cell spheroids [29][30][31] in contact with one another so they can fuse to form larger, more sophisticated tissue constructs. The Kenzan (Amuza, Inc., San Diego, CA) used was a 9 x 9 microneedle array made from stainless steel and housed in a biocompatible casing containing phosphate buffered saline.…”

Section: Biofabrication Equipment and The Kenzan Methodsmentioning

confidence: 99%

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Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

Wendel¹,

Wang²,

Smith³

et al. 2020

Preprint

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Endometriosis occurs when endometrial-like tissue grows outside the uterine cavity, leading to pelvic pain, infertility, and increased risk of ovarian cancer. The present study describes the optimization and characterization of cellular spheroids as building blocks for Kenzan scaffold-free method biofabrication and proof-of-concept models of endometriosis and the endometriotic microenvironment. The spheroid building blocks must be a specific diameter (~500 m), compact, round, and smooth to withstand Kenzan biofabrication. Under optimized spheroid conditions for biofabrication, the endometriotic epithelial-like cell line, 12Z, expressed high levels of estrogen-related genes and secreted high amounts of endometriotic inflammatory factors that were independent of TNF stimulation. Heterotypic spheroids, composed of 12Z and T-HESC, an immortalized endometrial stromal cell line, self-assembled into a biologically relevant pattern, consisting of epithelial cells on the outside of the spheroids and stromal cells in the core. 12Z spheroids were biofabricated into large three-dimensional constructs alone, with HEYA8 spheroids, or as heterotypic spheroids with T-HESC. These three-dimensional biofabricated constructs containing multiple monotypic or heterotypic spheroids represent the first scaffold-free biofabricated in vitro models of endometriosis and the endometriotic microenvironment. These efficient and innovative models will allow us to study the complex interactions of multiple cell types within a biologically relevant microenvironment.

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Section: Biofabrication Equipment and The Kenzan Methodsmentioning

confidence: 99%

Section: Biofabrication Equipment and The Kenzan Methodsmentioning

confidence: 99%

Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

Wendel¹,

Wang²,

Smith³

et al. 2020

Preprint

Self Cite

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“…The platen was aseptically coated with 0.15 mg/ml collagen (Rat Tail Collagen Type 1; Becton Dickenson Laboratories, Bedford, MA) for 1 hr, aspirating unbound collagen, and allowing the platen to air dry for at least 1 hr before use. This platen was then slotted onto a Kenzan needle array through its microchannels (Figure c,d) and then used for bioprinting tissue constructs from cellular spheroids into tissue constructs (Figures and ; Aguilar, Smith et al, ; Moldovan, Hibino, & Nakayama, ; Smith, Li, Holland, & Ekser, ).…”

Section: Methodsmentioning

confidence: 99%

“…Construct design was achieved using the Regenova (Cyfuse K.K., Japan) Bio 3D Printer's construct design feature. Following spheroid formation and construct design, the Regenova bioprinter was used to bioprint/biofabricate a scaffold‐free 3D tissue construct as described by Aguilar, Smith, et al () and Smith et al (). Briefly, Regenova's computer vision system selects a suitable spheroid and the bioprinter's mobile nozzle is then moved close to the spheroid and negative pneumatic back pressure is used to secure the spheroid to the nozzle tip and pick the selected spheroid from the 96‐well plate.…”

Section: Methodsmentioning

confidence: 99%

“…The scaffold‐dependent tissue is therefore dependent on these scaffolds to hold the tissue together over the long‐term. On the other hand, scaffold‐free is normally used to describe tissues that are comprised of cells embedded within their own cell‐secreted matrix (not contrived or reconstituted ECM scaffolds) and are therefore “scaffold‐free” (Aguilar, Olivos et al, ; Aguilar, Smith et al, ; Blakely, Manning, Tripathi, & Morgan, ; Itoh et al, ; Moldovan, ; Moldovan et al, ; Murata et al, ; Ozbolat, ; Toratani et al, ). The distinction between scaffold‐dependent and scaffold‐free can similarly be defined by the tissue's final composition after maturation.…”

Section: Methodsmentioning

confidence: 99%

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Computational fluid dynamic analysis of bioprinted self‐supporting perfused tissue models

Sego

Prideaux

Sterner

et al. 2019

Biotech & Bioengineering

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Natural tissues are incorporated with vasculature, which is further integrated with a cardiovascular system responsible for driving perfusion of nutrient-rich oxygenated blood through the vasculature to support cell metabolism within most cell-dense tissues. Since scaffold-free biofabricated tissues being developed into clinical implants, research models, and pharmaceutical testing platforms should similarly exhibit perfused tissue-like structures, we generated Abbreviations: CFD, computational fluid dynamics; ECM, extracellular matrix; FBS, fetal bovine serum; SSuPer, self-supporting perfused. 3D-bioprinting, biofabrication, bioreactor, computational fluid dynamics, perfusion, scaffold-free SUPPORTING INFORMATION Additional supporting information may be found online in the Supporting Information section. How to cite this article: Sego T, Prideaux M, Sterner J, et al. Computational fluid dynamic analysis of bioprinted self-supporting perfused tissue models. Biotechnology and Bioengineering. 2020;117:798-815.

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Converging 2D Nanomaterials and 3D Bioprinting Technology: State‐of‐the‐Art, Challenges, and Potential Outlook in Biomedical Applications

Rastin

Mansouri

Tùng

et al. 2021

Adv Healthcare Materials

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The development of next-generation of bioinks aims to fabricate anatomical size 3D scaffold with high printability and biocompatibility. Along with the progress in 3D bioprinting, 2D nanomaterials (2D NMs) prove to be emerging frontiers in the development of advanced materials owing to their extraordinary properties. Harnessing the properties of 2D NMs in 3D bioprinting technologies can revolutionize the development of bioinks by endowing new functionalities to the current bioinks. First the main contributions of 2D NMS in 3D bioprinting technologies are categorized here into six main classes: 1) reinforcement effect, 2) delivery of bioactive molecules, 3) improved electrical conductivity, 4) enhanced tissue formation, 5) photothermal effect, 6) and stronger antibacterial properties. Next, the recent advances in the use of each certain 2D NMs (1) graphene, 2) nanosilicate, 3) black phosphorus, 4) MXene, 5) transition metal dichalcogenides, 6) hexagonal boron nitride, and 7) metal-organic frameworks) in 3D bioprinting technology are critically summarized and evaluated thoroughly. Third, the role of physicochemical properties of 2D NMSs on their cytotoxicity is uncovered, with several representative examples of each studied 2D NMs. Finally, current challenges, opportunities, and outlook for the development of nanocomposite bioinks are discussed thoroughly.

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Scaffold-free bioprinting of mesenchymal stem cells with the regenova printer: Optimization of printing parameters

Cited by 38 publications

References 9 publications

Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

Three-Dimensional Biofabrication Models of Endometriosis and the Endometriotic Microenvironment

Computational fluid dynamic analysis of bioprinted self‐supporting perfused tissue models

Converging 2D Nanomaterials and 3D Bioprinting Technology: State‐of‐the‐Art, Challenges, and Potential Outlook in Biomedical Applications

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