可用于3D肝细胞培养的甘草次酸修饰海藻酸钠凝胶球的制备

doi:10.7503/cjcu20160316

高等学校化学学报 ›› 2017, Vol. 38 ›› Issue (2): 326.doi: 10.7503/cjcu20160316

• 高分子化学 • 上一篇

可用于3D肝细胞培养的甘草次酸修饰海藻酸钠凝胶球的制备

杨美跃¹, 王蔚¹(), 田志清¹, 李营营¹, 袁直^1,²()

1. 南开大学功能高分子教育部重点实验室, 高分子化学研究所, 天津 300071
2. 南开大学天津市化学化工协同创新中心, 天津 300071

收稿日期:2016-05-05 出版日期:2017-02-10 发布日期:2016-12-26
作者简介:联系人简介: 王蔚, 男, 博士, 副研究员, 主要从事生物医用材料研究. E-mail: duruo@nankai.edu.cn; 袁直, 女, 博士, 教授, 博士生导师, 主要从事生物医用材料研究. E-mail: zhiy@nankai.edu.cn
基金资助:
国家自然科学基金(批准号: 51273095)、天津市自然科学基金青年项目(批准号: 14JCQNJCO3500)、教育部长江学者创新团队发展计划项目(批准号: IRT1257)和国家基础科学人才培养基金(批准号: J1103306)资助

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

YANG Meiyue¹, WANG Wei^1,^*(), TIAN Zhiqing¹, LI Yingying¹, YUAN Zhi^1,^2,^*()

1. Key Laboratory of Functional Polymer Materials, Ministry of Education, Institute of Polymer Chemistry,Nakai University, Tianjin 300071, China
2. sourceorative Innovation Center of Chemical Science and Engineering(Tianjin),Nankai University, Tianjin 300071, China

Received:2016-05-05 Online:2017-02-10 Published:2016-12-26
Contact: WANG Wei,YUAN Zhi E-mail:duruo@nankai.edu.cn;zhiy@nankai.edu.cn
Supported by:
† Supported by the National Natural Science Foundation of China(No.51273095), the Natural Science Foundation of Tianjin, China(No.14JCQNJC03500), the Program for Changjiang Scholars and Innovative Research Team in University of Ministry of Education of China(No.IRT1257) and the National Found for Fostering Talents of Basic Science of China(No.J1103306)

摘要/Abstract

摘要：

制备了叔胺改性甘草次酸[GA-N(CH₃)₂]修饰的海藻酸钠[ALG-GA-N(CH₃)₂], 并在温敏性琼脂糖的辅助作用下, 利用微流体技术获得了高通量、单分散且粒径可控的ALG-GA-N(CH₃)₂微凝胶. 考察了Span 80含量、疏水配体取代度、样品浓度和水/油相流速对微液滴制备的影响. 研究结果表明, 叔胺基改性可显著改善甘草次酸的亲水性; 在Span 80质量分数为2.0%, 疏水配体取代度小于12%, 样品浓度小于15 mg/mL, 水相流速为1.5 mL/h, 油相流速为6 mL/h条件下, 可获得高通量、单分散及粒径为200 μm的适用于细胞包封培养的微凝胶球. 同时提供了一种三维培养肝细胞的新方法, 为其在组织工程中的应用奠定了基础.

关键词: 微流体, 海藻酸钠, 甘草次酸, 微凝胶球, 三维肝细胞培养

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

中图分类号:

O636
O648.17

TrendMD:

杨美跃, 王蔚, 田志清, 李营营, 袁直. 可用于3D肝细胞培养的甘草次酸修饰海藻酸钠凝胶球的制备. 高等学校化学学报, 2017, 38(2): 326.

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

图/表 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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可用于3D肝细胞培养的甘草次酸修饰海藻酸钠凝胶球的制备

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

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可用于3D肝细胞培养的甘草次酸修饰海藻酸钠凝胶球的制备

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

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