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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.


Schematic diagram showing the CD-CS method using a core–shell nozzle and single lines of cell-laden alginate.(a) The CD-CS cell-dispensing process with a nozzle with a 250-μm core and a 750-μm shell, where the cross-linking agent flowed in the shell and the cell-laden alginate flowed in the core. (b) A general process (GP) without cross-linking during dispensing. (c) The cell-dispensing process (CD-T) with an aerosol cross-linking process. (d) The CD-CS process. The schematic diagram was drawn by S.H. Ahn.
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f1: Schematic diagram showing the CD-CS method using a core–shell nozzle and single lines of cell-laden alginate.(a) The CD-CS cell-dispensing process with a nozzle with a 250-μm core and a 750-μm shell, where the cross-linking agent flowed in the shell and the cell-laden alginate flowed in the core. (b) A general process (GP) without cross-linking during dispensing. (c) The cell-dispensing process (CD-T) with an aerosol cross-linking process. (d) The CD-CS process. The schematic diagram was drawn by S.H. Ahn.

Mentions: We compared the stability of the of the cell-laden struts fabricated using three processes: a general process (GP) without cross-linking during the dispensing process15, a cell-dispensing process (CD-T) assisted via aerosol-based cross-linking1617, and a new cell-dispensing process (CD-CS) using a nozzle with a 250-μm core and a 750-μm shell, in which cross-linking agent flowed in the shell and a cell-laden hydrogel flowed in the core, as shown in Fig. 1(a). The pneumatic pressure of the mixture of 1 × 107 mL−1 of MC3T3-E1 cells in 3-wt% alginate was 70 kPa. With the CD-T process, the aerosol flow rate of the 5-wt% solution of CaCl2 was 1.4 mL min−1 and in the CD-CS process the flow rate of 1.2-wt% CaCl2 solution in the shell region was 0.08 mL min−1 18. To investigate the stability of the cylindrical shapes, surface and cross-sectional views of single-line cell-laden struts were obtained using an optical microscope. Figure 1(b–d) show optical microscope images of the single struts fabricated using the GP, CD-T and CD-CS processes. The single strut formed using the CD-CS process exhibited a completely round cross-section (in contrast to those formed using the GP and CD-T processes), which is attributed to the rapid gelation of the alginate immediately following making contact with the cross-linking agent. Based on this result, we may conclude the CD-CS process is suitable for the formation of cylindrical struts.


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)

Schematic diagram showing the CD-CS method using a core–shell nozzle and single lines of cell-laden alginate.(a) The CD-CS cell-dispensing process with a nozzle with a 250-μm core and a 750-μm shell, where the cross-linking agent flowed in the shell and the cell-laden alginate flowed in the core. (b) A general process (GP) without cross-linking during dispensing. (c) The cell-dispensing process (CD-T) with an aerosol cross-linking process. (d) The CD-CS process. The schematic diagram was drawn by S.H. Ahn.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Schematic diagram showing the CD-CS method using a core–shell nozzle and single lines of cell-laden alginate.(a) The CD-CS cell-dispensing process with a nozzle with a 250-μm core and a 750-μm shell, where the cross-linking agent flowed in the shell and the cell-laden alginate flowed in the core. (b) A general process (GP) without cross-linking during dispensing. (c) The cell-dispensing process (CD-T) with an aerosol cross-linking process. (d) The CD-CS process. The schematic diagram was drawn by S.H. Ahn.
Mentions: We compared the stability of the of the cell-laden struts fabricated using three processes: a general process (GP) without cross-linking during the dispensing process15, a cell-dispensing process (CD-T) assisted via aerosol-based cross-linking1617, and a new cell-dispensing process (CD-CS) using a nozzle with a 250-μm core and a 750-μm shell, in which cross-linking agent flowed in the shell and a cell-laden hydrogel flowed in the core, as shown in Fig. 1(a). The pneumatic pressure of the mixture of 1 × 107 mL−1 of MC3T3-E1 cells in 3-wt% alginate was 70 kPa. With the CD-T process, the aerosol flow rate of the 5-wt% solution of CaCl2 was 1.4 mL min−1 and in the CD-CS process the flow rate of 1.2-wt% CaCl2 solution in the shell region was 0.08 mL min−1 18. To investigate the stability of the cylindrical shapes, surface and cross-sectional views of single-line cell-laden struts were obtained using an optical microscope. Figure 1(b–d) show optical microscope images of the single struts fabricated using the GP, CD-T and CD-CS processes. The single strut formed using the CD-CS process exhibited a completely round cross-section (in contrast to those formed using the GP and CD-T processes), which is attributed to the rapid gelation of the alginate immediately following making contact with the cross-linking agent. Based on this result, we may conclude the CD-CS process is suitable for the formation of cylindrical struts.

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.