Effects of culturing media on hepatocytes differentiation using Volvox sphere as co-culturing vehicle

Kun Lieh Wu; Siou Han Chang; Ioannis Manousakas; Han Hsiang Huang; Benjamin Teong; Chin Wen Chuang; Shyh Ming Kuo

doi:10.1016/j.bbrc.2015.02.016

Effects of culturing media on hepatocytes differentiation using Volvox sphere as co-culturing vehicle

Biochem Biophys Res Commun. 2015 Mar 13;458(3):620-625. doi: 10.1016/j.bbrc.2015.02.016. Epub 2015 Feb 11.

Authors

Kun Lieh Wu¹, Siou Han Chang¹, Ioannis Manousakas¹, Han Hsiang Huang², Benjamin Teong¹, Chin Wen Chuang³, Shyh Ming Kuo⁴

Affiliations

¹ Department of Biomedical Engineering, I-Shou University, Kaohsiung City, Taiwan, ROC.
² Department of Veterinary Medicine, National Chiayi University, Chiayi City, Taiwan, ROC.
³ Department of Electric Engineering, I-Shou University, Kaohsiung City, Taiwan, ROC.
⁴ Department of Biomedical Engineering, I-Shou University, Kaohsiung City, Taiwan, ROC. Electronic address: smkuo@isu.edu.tw.

PMID: 25681769
DOI: 10.1016/j.bbrc.2015.02.016

Abstract

Volvox sphere is a unique design to mimic natural volvox consists of a large outer-sphere that contains smaller inner-spheres, which provide three-dimensional (3D) environment to culture cells. The purpose of this study is to co-culture mesenchymal stem cells (MSCs) and AML12 liver cells in Volvox spheres and to evaluate the effects of two media, DMEM and DMEM/F12 on the cultured cells. The results of this study shows that the 3D Volvox sphere can successfully be applied for co-culture of MSCs and AML12 liver cells, and the MSCs are able to differentiate into hepatocyte-like cells expressing hepatocyte-specific markers including albumin (ALB), alpha feto-protein (AFP) and cytokeratin 18 (CK18) mRNA expressions and producing CK18 and ALB proteins. Interestingly, the MSCs expressed higher ALB, AFP and CK18 mRNA expression at the initial 7-day culture by using DMEM, whereas, the MSCs expressed more mRNA expressions from 7-day to 14-day by the usage of DMEM/F12. The result demonstrated that DMEM and DMEM/F12 media could affect MSCs behaviors during a 14-day culture.

Keywords: Co-culture; Media; Mesenchymal stem cell; Volvox sphere.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Albumins / genetics
Animals
Biomimetic Materials* / chemistry
Cell Differentiation
Cell Line
Cells, Cultured
Cells, Immobilized / cytology
Cells, Immobilized / metabolism
Coculture Techniques / instrumentation
Coculture Techniques / methods*
Culture Media / metabolism*
Equipment Design
Hepatocytes / cytology*
Hepatocytes / metabolism
Keratin-18 / genetics
Mesenchymal Stem Cells / cytology*
Mesenchymal Stem Cells / metabolism
RNA, Messenger / genetics
Rats, Sprague-Dawley
Volvox* / chemistry
Volvox* / cytology
alpha-Fetoproteins / genetics

Substances

Albumins
Culture Media
Keratin-18
RNA, Messenger
alpha-Fetoproteins