A Simple Screening Method for Analysis of Cell Adhesive Strength Cultured on Zein Microspheres

doi:10.1007/s12204-017-1793-9

Abstract

Abstract: Cell adhesion plays an important role in the microcarrier culture. Therefore, it is necessary to find a simple method to evaluate the cell adhesive strength. By the glycerol method, we prepared zein microcarriers. Both microcarrier surface morphology and cells on the microcarriers could be observed clearly when the microcarriers and the cells were stained by acridine orange (AO). Pre-treatment of soaking and sterilizing processes was necessary for cell culture, but it did not alter the morphology of zein microcarriers. Three kinds of cells were cultured on zein microcarriers for 24 h. Vero cells showed the strongest adhesion, while L929 cells were easily detached from the microcarriers after a simple beating operation. The proliferation curve of Vero cells cultured on the microcarriers displayed no difference with that on Cytodex 1.

Key words: zein microcarrier| surface topography| cell adhesion| beating experiment

摘要： Cell adhesion plays an important role in the microcarrier culture. Therefore, it is necessary to find a simple method to evaluate the cell adhesive strength. By the glycerol method, we prepared zein microcarriers. Both microcarrier surface morphology and cells on the microcarriers could be observed clearly when the microcarriers and the cells were stained by acridine orange (AO). Pre-treatment of soaking and sterilizing processes was necessary for cell culture, but it did not alter the morphology of zein microcarriers. Three kinds of cells were cultured on zein microcarriers for 24 h. Vero cells showed the strongest adhesion, while L929 cells were easily detached from the microcarriers after a simple beating operation. The proliferation curve of Vero cells cultured on the microcarriers displayed no difference with that on Cytodex 1.

关键词: zein microcarrier| surface topography| cell adhesion| beating experiment

CLC Number:

Q 819

YANG Hui (杨辉), LI Weiying (李伟迎), WANG Jinye* (王瑾晔). A Simple Screening Method for Analysis of Cell Adhesive Strength Cultured on Zein Microspheres[J]. Journal of shanghai Jiaotong University (Science), 2017, 22(1): 10-014.

References 18

[1]	WU S C, HUANG G Y L. Stationary and microcarriercell culture processes for propagating Japaneseencephalitis virus [J]. Biotechnology Progress, 2002,18(1): 124-128.
[2]	ARIFIN M A, MEL M, KARIM M I A, et al. Productionof newcastle disease virus by Vero cells grown onCytodex 1 microcarriers in a 2-litre stirred tank bioreactor[J]. Journal of Biomedicine and Biotechnology,2010, 2010(1): 98-116.
[3]	CHU L L, ROBINSON D K. Industrial choices for proteinproduction by large-scale cell culture [J]. CurrentOpinion in Biotechnology, 2001, 12(2): 180-187.
[4]	ZHONG M L, SUN J, Wei D, et al. Establishinga cell-affinitive interface and spreading space in a3D hydrogel by introduction of microcarriers and anenzyme [J]. Jouranal of Materials Chemistry B, 2014,2: 6601-6610.
[5]	VARANI J, PIEL F, JOSEPHS S, et al. Attachmentand growth of anchorage-dependent cells on a novel,charged-surface microcarrier under serum-free conditions[J]. Cytotechnology, 1998, 28: 101-109.
[6]	HUPFELD J, GORR I H, SCHWALD C, et al. Modulationof mesenchymal stromal cell characteristics bymicrocarrier culture in bioreactors [J]. Biotechnologyand Bioengineering, 2014, 111(11): 2290-2302.
[7]	KINO Y, SAWA M, KASAI S, et al. Multiporous cellulosemicrocarrier for the development of a hybrid artificialliver using isolated hepatocytes [J]. Journal ofSurgical Research, 1998, 79(1): 71-76.
[8]	DONG J, SUN Q S, WANG J Y. Basic study of cornprotein, zein, as a biomaterial in tissue engineering,surface morphology and biocompatibility [J]. Biomaterials,2004, 25(19): 4691-4697.
[9]	LI W Y, HANG Y L, YANG H, et al. Preparation ofmicrocarriers based on zein and their application incell culture [J]. Materials Science and Engineering C,2016, 58: 863-869.
[10]	YEUNG T, GEORGES P C, FLANAGAN L A, etal. Effects of substrate stiffness on cell morphology,cytoskeletal structure and adhesion [J]. Cell Motilityand the Cytoskeleton, 2005, 60(1): 24-34.
[11]	CALIFANO J P, REINHART-KING C A. The effectsof substrate elasticity on endothelial cell network formationand traction force generation [C]//Proceedingsof 31st Annual International Conference of the IEEEEMBS. Minneapolis, USA: IEEE, 2009: 3343-3345.
[12]	LI B J, MOSHFEGH C, LIN Z, et al. Mesenchymalstem cells exploit extracellular matrix as mechanotransducer[J]. Scientific Reports, 2013, 3(33): 2425.
[13]	MARSHALL B T, LONG M, PIPER J W, et al. Directobservation of catch bonds involving cell-adhesionmolecules [J]. Nature, 2003, 423(6936): 190-193.
[14]	REYES C D, GARC′IAA J. A centrifugation cell adhesionassay for high-throughput screening of biomaterialsurfaces [J]. Journal of Biomedical Materials ResearchPart A, 2003, 67(1): 328-333.
[15]	MIER-TORRECILLA M, IDELSOHN S R, O?NATEE. Advances in the simulation of multi-fluid flows withthe particle finite element method: Application to bubbledynamics [J]. International Journal for NumericalMethods in Fluids, 2011, 67(11): 1516-1539.
[16]	ZHOU D W, GARC′IA A J. Measurement systems forcell adhesive forces [J]. Journal of Biomechanical Engineering,2015, 137(2): 020908.
[17]	BER S, K¨oSE G T, HASIRCI V. Bone tissue engineeringon patterned collagen films: An in vitro study [J].Biomaterials, 2005, 26(14): 1977-1986.
[18]	CHAREST J L, ELIASON M T, GARC′IA A J, etal. Combined microscale mechanical topography andchemical patterns on polymer substrates for cell culture[J]. Biomaterials, 2006, 27: 2487-2494.