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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.


Optimum conditions for the CD-CS process.(a) The stability of the cylindrical cell-laden alginates (3 and 5 wt%) for various calcium chloride solutions. (b) The cell-viability of the cell-laden struts formed with a 3-wt% concentration of alginate. Fluorescence images showing the cell viability for various CaCl2 solutions in shell nozzle, (c) 0.8, (d) 1.0, (e) 1.2, and (f) 1.4 wt%. In these images, the live cells are shown in green and dead cells shown in red.
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f4: Optimum conditions for the CD-CS process.(a) The stability of the cylindrical cell-laden alginates (3 and 5 wt%) for various calcium chloride solutions. (b) The cell-viability of the cell-laden struts formed with a 3-wt% concentration of alginate. Fluorescence images showing the cell viability for various CaCl2 solutions in shell nozzle, (c) 0.8, (d) 1.0, (e) 1.2, and (f) 1.4 wt%. In these images, the live cells are shown in green and dead cells shown in red.

Mentions: Figure 4(a) shows the effects of various mass factions of CaCl2 in the shell nozzle on the shape of the cell-laden alginate strut. A concentration of CaCl2 greater than 1.1 wt% in the shell region resulted in a stable cylindrical shape of the extruded struts, with b/a > 0.8. From this result, we may conclude that concentrations of CaCl2 greater than 1.1 wt% in the absorbent dispensing stage result in the formation of stable cylindrical cell-laden alginate struts, regardless of the limited mass fraction of the alginate-based bioink. The optical images shown in Fig. 4(a) reveal a stable cylindrical form of the dispensed bioink struts for solutions with a CaCl2 concentration greater than 1.2 wt%, whereas the cross-sectional image of the alginate strut, which was fabricated with a lower concentration of CaCl2 solution, did not maintain a stable cylindrical shape.


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)

Optimum conditions for the CD-CS process.(a) The stability of the cylindrical cell-laden alginates (3 and 5 wt%) for various calcium chloride solutions. (b) The cell-viability of the cell-laden struts formed with a 3-wt% concentration of alginate. Fluorescence images showing the cell viability for various CaCl2 solutions in shell nozzle, (c) 0.8, (d) 1.0, (e) 1.2, and (f) 1.4 wt%. In these images, the live cells are shown in green and dead cells shown in red.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Optimum conditions for the CD-CS process.(a) The stability of the cylindrical cell-laden alginates (3 and 5 wt%) for various calcium chloride solutions. (b) The cell-viability of the cell-laden struts formed with a 3-wt% concentration of alginate. Fluorescence images showing the cell viability for various CaCl2 solutions in shell nozzle, (c) 0.8, (d) 1.0, (e) 1.2, and (f) 1.4 wt%. In these images, the live cells are shown in green and dead cells shown in red.
Mentions: Figure 4(a) shows the effects of various mass factions of CaCl2 in the shell nozzle on the shape of the cell-laden alginate strut. A concentration of CaCl2 greater than 1.1 wt% in the shell region resulted in a stable cylindrical shape of the extruded struts, with b/a > 0.8. From this result, we may conclude that concentrations of CaCl2 greater than 1.1 wt% in the absorbent dispensing stage result in the formation of stable cylindrical cell-laden alginate struts, regardless of the limited mass fraction of the alginate-based bioink. The optical images shown in Fig. 4(a) reveal a stable cylindrical form of the dispensed bioink struts for solutions with a CaCl2 concentration greater than 1.2 wt%, whereas the cross-sectional image of the alginate strut, which was fabricated with a lower concentration of CaCl2 solution, did not maintain a stable cylindrical shape.

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.