3D Bioprinting of Human Adipose-Derived Stem Cells and Their Tenogenic Differentiation in Clinical-Grade Medium

Defining the best combination of cells and biomaterials is a key challenge for the development of tendon tissue engineering (TE) strategies. Adipose-derived stem cells (ASCs) are ideal candidates for this purpose. In addition, controlled cell-based products adherent to good manufacturing practice (GMP) are required for their clinical scale-up. With this aim, in this study, ASC 3D bioprinting and GMP-compliant tenogenic differentiation were investigated. In detail, primary human ASCs were embedded within a nanofibrillar-cellulose/alginate bioink and 3D-bioprinted into multi-layered square-grid matrices. Bioink viscoelastic properties and scaffold ultrastructural morphology were analyzed by rheology and scanning electron microscopy (SEM). The optimal cell concentration for printing among 3, 6 and 9 × 106 ASC/mL was evaluated in terms of cell viability. ASC morphology was characterized by SEM and F-actin immunostaining. Tenogenic differentiation ability was then evaluated in terms of cell viability, morphology and expression of scleraxis and collagen type III by biochemical induction using BMP-12, TGF-β3, CTGF and ascorbic acid supplementation (TENO). Pro-inflammatory cytokine release was also assessed. Bioprinted ASCs showed high viability and survival and exhibited a tenocyte-like phenotype after biochemical induction, with no inflammatory response to the bioink. In conclusion, we report a first proof of concept for the clinical scale-up of ASC 3D bioprinting for tendon TE.

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Scale‐up of an intensified bioprocess for the expansion of bovine adipose‐derived stem cells (bASCs) in stirred tank bioreactors

Biotechnology and Bioengineering ◽

10.1002/bit.27842 ◽

2021 ◽

Author(s):

Mariana Petronela Hanga ◽

Fritz Anthony Raga ◽

Panagiota Moutsatsou ◽

Christopher J. Hewitt ◽

Alvin W. Nienow ◽

...

Keyword(s):

Stem Cells ◽

Scale Up ◽

Stirred Tank ◽

Adipose Derived Stem Cells

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Tendon Tissue Engineering: Mechanism and Effects of Human Tenocyte Coculture With Adipose-Derived Stem Cells

The Journal Of Hand Surgery ◽

10.1016/j.jhsa.2017.07.031 ◽

2018 ◽

Vol 43 (2) ◽

pp. 183.e1-183.e9 ◽

Cited By ~ 5

Author(s):

Chao Long ◽

Zhen Wang ◽

Anais Legrand ◽

Arhana Chattopadhyay ◽

James Chang ◽

...

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Adipose Derived Stem Cells ◽

Tendon Tissue ◽

Tendon Tissue Engineering

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Effect of Uniaxial Tensile Cyclic Loading Regimes on Matrix Organization and Tenogenic Differentiation of Adipose-Derived Stem Cells Encapsulated within 3D Collagen Scaffolds

Stem Cells International ◽

10.1155/2017/6072406 ◽

2017 ◽

Vol 2017 ◽

pp. 1-16 ◽

Cited By ~ 5

Author(s):

Gayathri Subramanian ◽

Alexander Stasuk ◽

Mostafa Elsaadany ◽

Eda Yildirim-Ayan

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Expression Profiles ◽

Three Dimensional ◽

Uniaxial Tensile ◽

Adipose Derived Stem Cells ◽

Collagen Scaffolds ◽

Differentiation Potential ◽

Tenogenic Differentiation ◽

Matrix Organization

Adipose-derived mesenchymal stem cells have become a popular cell choice for tendon repair strategies due to their relative abundance, ease of isolation, and ability to differentiate into tenocytes. In this study, we investigated the solo effect of different uniaxial tensile strains and loading frequencies on the matrix directionality and tenogenic differentiation of adipose-derived stem cells encapsulated within three-dimensional collagen scaffolds. Samples loaded at 0%, 2%, 4%, and 6% strains and 0.1 Hz and 1 Hz frequencies for 2 hours/day over a 7-day period using a custom-built uniaxial tensile strain bioreactor were characterized in terms of matrix organization, cell viability, and musculoskeletal gene expression profiles. The results displayed that the collagen fibers of the loaded samples exhibited increased matrix directionality with an increase in strain values. Gene expression analyses demonstrated that ASC-encapsulated collagen scaffolds loaded at 2% strain and 0.1 Hz frequency showed significant increases in extracellular matrix genes and tenogenic differentiation markers. Importantly, no cross-differentiation potential to osteogenic, chondrogenic, and myogenic lineages was observed at 2% strain and 0.1 Hz frequency loading condition. Thus, 2% strain and 0.1 Hz frequency were identified as the appropriate mechanical loading regime to induce tenogenic differentiation of adipose-derived stem cells cultured in a three-dimensional environment.

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Electromagnetic fields enhance chondrogenesis of human adipose-derived stem cells in a chondrogenic microenvironment in vitro

Journal of Applied Physiology ◽

10.1152/japplphysiol.01216.2012 ◽

2013 ◽

Vol 114 (5) ◽

pp. 647-655 ◽

Cited By ~ 33

Author(s):

Chung-Hwan Chen ◽

Yi-Shan Lin ◽

Yin-Chih Fu ◽

Chih-Kuang Wang ◽

Shun-Cheng Wu ◽

...

Keyword(s):

Stem Cells ◽

Electromagnetic Field ◽

Cell Viability ◽

Chondrogenic Differentiation ◽

Collagen Type ◽

Cartilage Tissue ◽

Pulse Electromagnetic Field ◽

Type I ◽

Adipose Derived Stem Cells ◽

Pemf Treatment

We tested the hypothesis that electromagnetic field (EMF) stimulation enhances chondrogenesis in human adipose-derived stem cells (ADSCs) in a chondrogenic microenvironment. A two-dimensional hyaluronan (HA)-coated well (2D-HA) and a three-dimensional pellet culture system (3D-pellet) were used as chondrogenic microenvironments. The ADSCs were cultured in 2D-HA or 3D-pellet, and then treated with clinical-use pulse electromagnetic field (PEMF) or the innovative single-pulse electromagnetic field (SPEMF) stimulation. The cytotoxicity, cell viability, and chondrogenic and osteogenic differentiations were analyzed after PEMF or SPEMF treatment. The modules of PEMF and SPEMF stimulations used in this study did not cause cytotoxicity or alter cell viability in ADSCs. Both PEMF and SPEMF enhanced the chondrogenic gene expression (SOX-9, collagen type II, and aggrecan) of ADSCs cultured in 2D-HA and 3D-pellet. The expressions of bone matrix genes (osteocalcin and collagen type I) of ADSCs were not changed after SPEMF treatment in 2D-HA and 3D-pellet; however, they were enhanced by PEMF treatment. Both PEMF and SPEMF increased the cartilaginous matrix (sulfated glycosaminoglycan) deposition of ADSCs. However, PEMF treatment also increased mineralization of ADSCs, but SPEMF treatment did not. Both PEMF and SPEMF enhanced chondrogenic differentiation of ADSCs cultured in a chondrogenic microenvironment. SPEMF treatment enhanced ADSC chondrogenesis, but not osteogenesis, when the cells were cultured in a chondrogenic microenvironment. However, PEMF enhanced both osteogenesis and chondrogenesis under the same conditions. Thus the combination of a chondrogenic microenvironment with SPEMF stimulation can promote chondrogenic differentiation of ADSCs and may be applicable to articular cartilage tissue engineering.

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Tuning the Thermogelation and Rheology of Poly(2-Oxazoline)/poly(2-Oxazine)s Based Thermosensitive Hydrogels for 3D Bioprinting.

10.26434/chemrxiv.14472114.v1 ◽

2021 ◽

Author(s):

Malik Salman Haider ◽

Taufiq Ahmad ◽

Mengshi Yang ◽

Chen Hu ◽

Lukas Hahn ◽

...

Keyword(s):

Drug Delivery ◽

Stem Cells ◽

Gel Strength ◽

Adipose Derived Stem Cells ◽

3D Bioprinting ◽

Thermosensitive Hydrogel ◽

Clay Nanoparticles ◽

Hydrogel Matrix ◽

Thermosensitive Hydrogels ◽

3D Printed

As one kind of smart material, thermogelling polymers find applications in biofabrication, drug delivery and regenerative medicine. Here, we reported on a novel thermosensitive hydrogel which can be 3D printed using extrusion based printing. Gel strength was found around 3kPa storage modulus with pronounced shear thinning and rapid recovery after stress. Addition of clay nanoparticles (Laponite XLG) improved the rheological profile further. Human adipose derived stem cells were added to the hydrogel matrix, which remained fully viable after printing. Therefore, the presented materials adds to the available material toolbox for 3D bioprinting. <br>

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