Preparation of Glycyrrhetinic Acid-modified Sodium Alginate Microgel Spheres for 3D Cell Culture†

doi:10.7503/cjcu20160316

Abstract

Abstract:

In order to obtain microgel sphere containing hydrophobic ligands which can be used for 3D liver cell culture. Firstly, the sodium alginate(ALG) was modified with tertiary ammonia-glycyrrhetinic acid[GA-N(CH₃)₂]. Then, the high flux, monodisperse, size controllable microgel of GA-N(CH₃)₂ modified ALG[ALG-GA-N(CH₃)₂] was prepared by the technology of microfluidics with the assistant of agarose. The influence of different preparation condition, such as concentration of Span 80, substitution degree of hydrophobic ligand, concentration of ALG-GA-N(CH₃)₂, velocity of water/oil phase, on microdroplet preparation were also studied. The results showed that the introduction of tertiary ammonia can significantly improve the hydrophility of GA. And the monodisperse microgel with 200 μm diameter, which could be used for cell envelope culture, could be obtained under the condition below: the mass fraction of Span 80 is 2.0%, hydrophobic ligand substitution degree is less than 12%, concentration of sample is less than 15 mg/mL, the velocity of water is 1.5 mL/h and the velocity of oil is 6.0 mL/h. The microgel provides a new method for liver cell 3D culture and laid the foundation for its application in tissue engineering.

Key words: Microfluidics, Sodium alginate, Glycyrrhetinic acid, Microgel sphere, 3D liver cell culture

CLC Number:

O636
O648.17

TrendMD:

YANG Meiyue, WANG Wei, TIAN Zhiqing, LI Yingying, YUAN Zhi. Preparation of Glycyrrhetinic Acid-modified Sodium Alginate Microgel Spheres for 3D Cell Culture^†[J]. Chem. J. Chinese Universities, 2017, 38(2): 326.

Figures/Tables 11

Scheme 1 Synthesis routes of en-GA-N(CH3)2(4)

Scheme 2 Synthesis route of ALG-GA-N(CH3)2

Fig.1 1H NMR spectra of GA(a), GA-N(CH3)2(b), suc-GA-N(CH3)2(c) and en-GA-N(CH3)2(d)

Fig.2 FTIR spectra of ALG(a) and ALG-GA-N(CH3)2(b)

Fig.3 Contact angles of water drop on glass(A), GA(B) and GA-N(CH3)2(C) surfaces Contact angle/(°): (A) 24.2; (B) 75.4; (C) 57.0.

Fig.4 Fluorescence curves of pyrene solution with different ALG-GA-N(CH3)2 concentration(A) and CAC value of ALG-GA-N(CH3)2(B)DS=5.68%. c[ALG-GA-N(CH3)2]/(mg·mL-1): a. 0.001; b. 0.05; c. 0.07; d. 0.09; e. 0.15; f. 0.25; g. 1.

Fig.5 Optical images of droplets formation with different Span 80 concentration voil=3.0 mL/h, vwater=1.5 mL/h. Mass fraction of Span 80(%): (A) 0; (B) 2; (C) 5; (D) 8.

Fig.6 Influence of flow rate of oil phase(A) and water phase(B) on the diameter of droplets (A) v(Water)=0.5 mL/h; (B) v(Oil)=1.3 mL/h.

Fig.7 Optical images of droplets formation of ALG-GA-N(CH3)2 with different DS of GA-N(CH3)2 c[ALG-GA-N(CH3)2]=12 mg/mL; v(Water)=1.5 mL/h, v(Oil)=6.0 mL/h. (A) DS=4.3%; (B) DS=8.6%; (C) DS=12.0%.

Fig.8 Optical images of droplets formation of ALG-GA-N(CH3)2 with different concentration DS=8.6%; v(Water)=1.5 mL/h, v(Oil)=6.0 mL/h. Concentration of ALG-GA-N(CH3)2/(mg·mL-1): (A) 9; (B) 12; (C) 15; (D) 18.

Fig.9 Optical images of ALG-GA-N(CH3)2 microgel spheres

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