淀粉微载体的制备及对CHO-K1细胞培养的初步应用

doi:10.16333/j.1001-6880.2017.3.017

天然产物研究与开发 ›› 2017, Vol. 29 ›› Issue (3): 454-460.doi: 10.16333/j.1001-6880.2017.3.017

淀粉微载体的制备及对CHO-K1细胞培养的初步应用

张磊1,2，李明生2,3，郑荣1，张健1，靳冬武1，马忠仁1,2，冯玉萍1,2*

1 西北民族大学生命科学与工程学院；2 西北民族大学生物工程与技术国家民委重点实验室；3 甘肃省动物细胞工程技术研究中心，兰州 730030

出版日期:2017-03-31 发布日期:2017-04-06

Preparation of Starch Microcarriers and Its Application in CHO-K1 Cell Culture

ZHANG Lei1,2,LI Ming-sheng2,3,ZHENG Rong1, ZHANG Jian1,JIN Dong-wu1,MA Zhong-ren1,2,FENG Yu-ping1,2*

1 Life Science and Engineering College of Northwest University for Nationalities;2Key  Laboratory of Bioengineering & Biotechnology of State Ethnic Affair Commission,Northwest University for Nationalities; 3Engineering Technology Research Center for Animal Cell,Lanzhou 730030,China

Online:2017-03-31 Published:2017-04-06

摘要/Abstract

摘要： 以普通市售马铃薯淀粉为原材料，在常温下利用W/O乳化方法制备了交联淀粉微球，并以二乙胺基乙基修饰了微球的表面电荷。同时研究了制备淀粉微球的制备工艺和改性工艺，成功制备了适合细胞培养用的淀粉微载体。利用扫描电镜（SEM）、红外（FT-IR）、X射线衍射仪（XRD）对微球进行了表征。结果表明，最佳制备条件为：淀粉溶液浓度为7%，搅拌速度为500 rpm，交联剂用量为8%，水油相体积比1:6；微球表面修饰最佳条件为：加入微球质量2倍体积的3.5 mol/L NaOH溶液和2.5 mol/L DEAE-HCl溶液，60 ℃密闭反应4 h。对比Cytodex-1微载体，培养CHO-K1细胞至144 h时，淀粉微载体表面细胞密度达到1.8×10⁶ cells/mL，且两者培养效果相似，表明了自制淀粉微载体是一种潜在的贴壁细胞培养用高分子材料。

关键词: 马铃薯淀粉, 淀粉微球, 二乙氨基乙基, CHO-K1细胞, 细胞培养

Abstract: The cross-linked starch microsphere(PCSM) was synthesized from potato starch by W/O emulsion method at room temperature,while its surface charge was modified with 2-dimethylaminoethyl.Meanwhile,the preparation and modification processes were studied,and the suitable starch microcarrier was successfully made for cell cultivation.The microspheres were characterized by scanning electron microscopy(SEM),Fourier translation infrared spectrum(FTIR) and X-ray diffraction(XRD).The results showed that the optimal parameters for the preparation were as follows:the starch solution concentration of 7%,the mixing speed of 500 RPM,the crosslinking agent dosage of 8%,and the oil/water volume proportion of 1:6.The optimal parameters for the modification process were found:2 times the mass of microspheres in volume of 3.5 mol/L NaOH solution and 2.5 mol/L DEAE-HCl solution added,and then stirred at 60 ℃ for 4 h.Compared with Cytodex-1,after culturing CHO-K1 for 144 h,the cell density on the surface of starch microcarrier reached 1.8 x 10⁶ cells/mL.The culturing effect remained similar for both microcarrier,suggesting that the homemade starch microcarrier is a kind of potential high polymer material for adherent cell culture.

Key words: potato starch, starch microspheres, 2-dimethylaminoethyl, CHO-K1 cells, cell culture

中图分类号:

Q813.1

张磊1,2，李明生2,3，郑荣1，张健1，靳冬武1，马忠仁1,2，冯玉萍1,2*. 淀粉微载体的制备及对CHO-K1细胞培养的初步应用[J]. 天然产物研究与开发, 2017, 29(3): 454-460.

ZHANG Lei1,2,LI Ming-sheng2,3,ZHENG Rong1, ZHANG Jian1,JIN Dong-wu1,MA Zhong-ren1,2,FENG Yu-ping1,2*. Preparation of Starch Microcarriers and Its Application in CHO-K1 Cell Culture[J]. NATURAL PRODUCT RESEARCH AND DEVELOPMENT, 2017, 29(3): 454-460.

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淀粉微载体的制备及对CHO-K1细胞培养的初步应用

Preparation of Starch Microcarriers and Its Application in CHO-K1 Cell Culture

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