Cost-Effective Production of Biologically Active Leukemia Inhibitory Factor for Mouse Embryonic Stem Cell Culture.
Molecular biotechnology, vol.68, no.4, pp.1825-1836, 2026 (SCI-Expanded, Scopus)
- Publication Type: Article / Article
- Volume: 68 Issue: 4
- Publication Date: 2026
- Doi Number: 10.1007/s12033-025-01478-6
- Journal Name: Molecular biotechnology
- Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Biotechnology Research Abstracts, CAB Abstracts, Chemical Abstracts Core, Compendex, Food Science & Technology Abstracts, Index Islamicus, INSPEC, MEDLINE, Veterinary Science Database
- Page Numbers: pp.1825-1836
- Keywords: Recombinant HM-LIF, Mouse embryonic stem cell, Affinity chromatography, Pluripotency, Neuronal differentiation
- Dokuz Eylül University Affiliated: Yes
Abstract
Leukemia inhibitory factor (LIF), a glycoprotein in the interleukin-6 family, is essential for maintaining the pluripotency and self-renewal of mouse embryonic stem cells (mESCs) by activating the JAK/STAT pathway and preventing spontaneous differentiation. Despite its critical role in maintaining the undifferentiated state of ES cells, the high cost of commercially available LIF poses a considerable financial challenge for research laboratories. This financial strain can limit the accessibility and feasibility of high-quality stem cell research, thereby impacting the overall progress in this field. Here, we present an efficient and cost-effective method for recombinant expression and purification of biologically active, non-glycosylated mouse LIF in Escherichia coli, which retains full functionality in mESC culture. Utilizing an N-terminal glutathione S-transferase (GST) tag enhances the solubility of LIF, facilitating rapid purification via glutathione-agarose affinity chromatography. Following purification, LIF is cleaved from the GST tag using HRV 3C protease, resulting in a product ready for direct application in mESC culture. Homemade LIF (HM-LIF) prepared with this protocol effectively maintains mESCs in an undifferentiated state, showing comparable efficacy to commercial LIF. Furthermore, mESCs cultured with HM-LIF retain pluripotency and can differentiate into glutamatergic neurons. This streamlined approach significantly reduces the cost of mESC culture while providing a reliable, high-quality alternative to commercial LIF.