Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture

Sumeth Imsoonthornruksa; Kamthorn Pruksananonda; Rangsun Parnpai; Ruttachuk Rungsiwiwut; Mariena Ketudat-Cairns

doi:10.1159/000441453

Expression and Purification of Recombinant Human Basic Fibroblast Growth Factor Fusion Proteins and Their Uses in Human Stem Cell Culture

J Mol Microbiol Biotechnol. 2015;25(6):372-80. doi: 10.1159/000441453. Epub 2015 Nov 20.

Authors

Sumeth Imsoonthornruksa¹, Kamthorn Pruksananonda, Rangsun Parnpai, Ruttachuk Rungsiwiwut, Mariena Ketudat-Cairns

Affiliation

¹ Embryo Technology and Stem Cell Research Center and School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima, Thailand.

PMID: 26584430
DOI: 10.1159/000441453

Abstract

To reduce the cost of cytokines and growth factors in stem cell research, a simple method for the production of soluble and biological active human basic fibroblast growth factor (hbFGF) fusion protein in Escherichia coli was established. Under optimal conditions, approximately 60-80 mg of >95% pure hbFGF fusion proteins (Trx-6xHis-hbFGF and 6xHis-hbFGF) were obtained from 1 liter of culture broth. The purified hbFGF proteins, both with and without the fusion tags, were biologically active, which was confirmed by their ability to stimulate proliferation of NIH3T3 cells. The fusion proteins also have the ability to support several culture passages of undifferentiated human embryonic stem cells and induce pluripotent stem cells. This paper describes a low-cost and uncomplicated method for the production and purification of biologically active hbFGF fusion proteins.

Publication types

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

MeSH terms

Alkaline Phosphatase / metabolism
Animals
Cell Culture Techniques / economics
Cell Culture Techniques / methods*
Embryonic Stem Cells / cytology*
Embryonic Stem Cells / drug effects
Escherichia coli / genetics
Escherichia coli / metabolism
Fibroblast Growth Factor 2 / biosynthesis*
Fibroblast Growth Factor 2 / genetics
Fibroblast Growth Factor 2 / isolation & purification*
Fibroblast Growth Factor 2 / pharmacology
Gene Expression Regulation, Bacterial
Genetic Vectors
Humans
Immunohistochemistry
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / drug effects
Mice
NIH 3T3 Cells
Polymerase Chain Reaction
Protein Engineering
Recombinant Fusion Proteins / biosynthesis*
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / isolation & purification*
Recombinant Fusion Proteins / pharmacology
Recombinant Proteins / biosynthesis*
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification*
Recombinant Proteins / pharmacology

Substances

KCB-1 protein, recombinant
Recombinant Fusion Proteins
Recombinant Proteins
Fibroblast Growth Factor 2
Alkaline Phosphatase