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A novel cell-printing method and its application to hepatogenic differentiation of human adipose stem cell-embedded mesh structures.

Ahn SH, Lee HJ, Lee JS, Yoon H, Chun W, Kim GH - Sci Rep (2015)

Bottom Line: In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a three-dimensional mesh structure.To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures.The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomechatronic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University (SKKU), Suwon, South Korea.

ABSTRACT
We report a cell-dispensing technique, using a core-shell nozzle and an absorbent dispensing stage to form cell-embedded struts. In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a three-dimensional mesh structure. The cell-printing conditions were optimized by manipulating the process conditions to obtain high mechanical stability and high cell viability. The cell density was 1 × 10(7) mL(-1), which was achieved using a 3-wt% solution of alginate in phosphate-buffered saline, a mass fraction of 1.2 wt% of CaCl2 flowing in the shell nozzle with a fixed flow rate of 0.08 mL min(-1), and a translation velocity of the printing nozzle of 10 mm s(-1). To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures. The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.

No MeSH data available.


Optical and SEM images and cellular activities of the 3D cell-laden mesh structures.Optical and SEM images of 3D MC3T3-E1-laden mesh structures fabricated using CD-CS. (a) 3D view, (b) surface view, and (c) side view. (d) Fluorescence images showing the cell viability of the cell-laden mesh structures fabricated using the CD-T and CD-CS after 1 day. (e) Cell viability measured using the fluorescence images and (f) MTT assay results showing cell proliferation. (g) Relative expression of ALP, BMP-2, OCN, and Col-I gene expression levels in MC3T3-E1 cells for 14 days. In the fluorescence images, the live cells are shown in green and dead cells shown in red. Asterisks (*) indicate p < 0.05.
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f5: Optical and SEM images and cellular activities of the 3D cell-laden mesh structures.Optical and SEM images of 3D MC3T3-E1-laden mesh structures fabricated using CD-CS. (a) 3D view, (b) surface view, and (c) side view. (d) Fluorescence images showing the cell viability of the cell-laden mesh structures fabricated using the CD-T and CD-CS after 1 day. (e) Cell viability measured using the fluorescence images and (f) MTT assay results showing cell proliferation. (g) Relative expression of ALP, BMP-2, OCN, and Col-I gene expression levels in MC3T3-E1 cells for 14 days. In the fluorescence images, the live cells are shown in green and dead cells shown in red. Asterisks (*) indicate p < 0.05.

Mentions: To obtain the 3D MC3T3-E1-laden mesh structures, stable processing conditions (i.e., a cell density of 1 × 107 mL−1, a 3-wt% alginate solution in PBS, a pneumatic pressure of 70 kPa in the core region of the nozzle, a CaCl2 flow rate of 0.08 mL min−1 in shell region of the nozzle, and translation speed of 10 mm s−1) were used with CD-CS with the absorbent stage. Figure 5(a–c) show optical microscope and scanning electron microscope (SEM) images of the fabricated cell-laden multi-layered mesh-structure (with dimensions of 20 × 20 × 5 mm3) in which the strut diameter was 414 ± 39 μm and pore size between the struts was 527 ± 28 μm.


A novel cell-printing method and its application to hepatogenic differentiation of human adipose stem cell-embedded mesh structures.

Ahn SH, Lee HJ, Lee JS, Yoon H, Chun W, Kim GH - Sci Rep (2015)

Optical and SEM images and cellular activities of the 3D cell-laden mesh structures.Optical and SEM images of 3D MC3T3-E1-laden mesh structures fabricated using CD-CS. (a) 3D view, (b) surface view, and (c) side view. (d) Fluorescence images showing the cell viability of the cell-laden mesh structures fabricated using the CD-T and CD-CS after 1 day. (e) Cell viability measured using the fluorescence images and (f) MTT assay results showing cell proliferation. (g) Relative expression of ALP, BMP-2, OCN, and Col-I gene expression levels in MC3T3-E1 cells for 14 days. In the fluorescence images, the live cells are shown in green and dead cells shown in red. Asterisks (*) indicate p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4544022&req=5

f5: Optical and SEM images and cellular activities of the 3D cell-laden mesh structures.Optical and SEM images of 3D MC3T3-E1-laden mesh structures fabricated using CD-CS. (a) 3D view, (b) surface view, and (c) side view. (d) Fluorescence images showing the cell viability of the cell-laden mesh structures fabricated using the CD-T and CD-CS after 1 day. (e) Cell viability measured using the fluorescence images and (f) MTT assay results showing cell proliferation. (g) Relative expression of ALP, BMP-2, OCN, and Col-I gene expression levels in MC3T3-E1 cells for 14 days. In the fluorescence images, the live cells are shown in green and dead cells shown in red. Asterisks (*) indicate p < 0.05.
Mentions: To obtain the 3D MC3T3-E1-laden mesh structures, stable processing conditions (i.e., a cell density of 1 × 107 mL−1, a 3-wt% alginate solution in PBS, a pneumatic pressure of 70 kPa in the core region of the nozzle, a CaCl2 flow rate of 0.08 mL min−1 in shell region of the nozzle, and translation speed of 10 mm s−1) were used with CD-CS with the absorbent stage. Figure 5(a–c) show optical microscope and scanning electron microscope (SEM) images of the fabricated cell-laden multi-layered mesh-structure (with dimensions of 20 × 20 × 5 mm3) in which the strut diameter was 414 ± 39 μm and pore size between the struts was 527 ± 28 μm.

Bottom Line: In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a three-dimensional mesh structure.To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures.The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomechatronic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University (SKKU), Suwon, South Korea.

ABSTRACT
We report a cell-dispensing technique, using a core-shell nozzle and an absorbent dispensing stage to form cell-embedded struts. In the shell of the nozzle, a cross-linking agent flowed continuously onto the surface of the dispensed bioink in the core nozzle, so that the bioink struts were rapidly gelled, and any remnant cross-linking solution during the process was rapidly absorbed into the working stage, resulting in high cell-viability in the bioink strut and stable formation of a three-dimensional mesh structure. The cell-printing conditions were optimized by manipulating the process conditions to obtain high mechanical stability and high cell viability. The cell density was 1 × 10(7) mL(-1), which was achieved using a 3-wt% solution of alginate in phosphate-buffered saline, a mass fraction of 1.2 wt% of CaCl2 flowing in the shell nozzle with a fixed flow rate of 0.08 mL min(-1), and a translation velocity of the printing nozzle of 10 mm s(-1). To demonstrate the applicability of the technique, preosteoblasts and human adipose stem cells (hASCs) were used to obtain cell-laden structures with multi-layer porous mesh structures. The fabricated cell-laden mesh structures exhibited reasonable initial cell viabilities for preosteoblasts (93%) and hASCs (92%), and hepatogenic differentiation of hASC was successfully achieved.

No MeSH data available.