PROKARYOTIC EXPRESSION AND PURIFICATION OF RECOMBINANT HUMANLEUKEMIA INHIBITORY FACTOR; ANALYSIS OF THE ABILITY TO MAINTAINPLURIPOTENCY IN EMBRYONIC STEM CELLS

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TAHERI BEHROUZ; MODARRESSI MOHAMMAD HOSSEIN; HASHEMITABAR MAHMOUD; MIRYOUNESI MOHAMMAD; MOTEVASELI ELAHE

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Journal: Iranian Red Crescent Medical Journal Year:2018 | Volume:20 | Issue:8 Page(s): 0-0

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Title

PROKARYOTIC EXPRESSION AND PURIFICATION OF RECOMBINANT HUMANLEUKEMIA INHIBITORY FACTOR; ANALYSIS OF THE ABILITY TO MAINTAINPLURIPOTENCY IN EMBRYONIC STEM CELLS

Author(s)

Keywords

EMBRYONIC STEM CELL

LEUKEMIA INHIBITORY FACTOR

PLURIPOTENCY

RECOMBINANT PROTEIN

Abstract

Background: LEUKEMIA INHIBITORY FACTOR (LIF) is largely used in stem cell researches for the maintenance of EMBRYONIC STEM CELLs(ESCs) in a pluripotent state. However, the relatively high costs of LIF is a potential limit of such researches. Objectives: The aim of this study was prokaryotic expression and purification of the recombinant hexahistidine-tagged human LIF(His6-hLIF) fusion protein and assessment of its ability to maintain a pluripotent or undifferentiated state of ESCs. Methods: Encoding the DNA sequence of mature hLIF was codon optimized for expression in Escherichia coli and chemically synthesizedand cloned in the expression vector pET-28a (+). Immobilized Metal Affinity Chromatography (IMAC) was performed topurify the recombinant His6-hLIF. Then, His6-hLIF was tested for its ability to maintain mESC by comparison with commercial LIF asa control. Results: The yield for the recombinant His6-hLIF was assessed to be approximately 1. 7 mg from one liter of culture. There were nostatistically significant differences in expression of two PLURIPOTENCY gene markers, oct-4 and Nanog, between mESCs treated withHis6-hLIF and those with commercial hLIF (P = 0. 09 and P = 0. 13, respectively). Besides, morphological characteristics (round cellularmorphology) were similar between them. Conclusions: Collectively, the findings showed that the ability of the recombinant His6-hLIF protein in maintaining pluripotentstate of ESCs was comparable to commercial hLIF, providing evidence that the presence of the N-terminal hexahistidin tag does notinfluence biological activities of hLIF.

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References

Expression, purification and characterization of active untagged recombinant human leukemia inhibitory factor from E. coli.

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APA: Copy

TAHERI, BEHROUZ, MODARRESSI, MOHAMMAD HOSSEIN, HASHEMITABAR, MAHMOUD, MIRYOUNESI, MOHAMMAD, & MOTEVASELI, ELAHE. (2018). PROKARYOTIC EXPRESSION AND PURIFICATION OF RECOMBINANT HUMANLEUKEMIA INHIBITORY FACTOR; ANALYSIS OF THE ABILITY TO MAINTAINPLURIPOTENCY IN EMBRYONIC STEM CELLS. IRANIAN RED CRESCENT MEDICAL JOURNAL (IRCMJ), 20(8), 0-0. SID. https://sid.ir/paper/293794/en

Vancouver: Copy

TAHERI BEHROUZ, MODARRESSI MOHAMMAD HOSSEIN, HASHEMITABAR MAHMOUD, MIRYOUNESI MOHAMMAD, MOTEVASELI ELAHE. PROKARYOTIC EXPRESSION AND PURIFICATION OF RECOMBINANT HUMANLEUKEMIA INHIBITORY FACTOR; ANALYSIS OF THE ABILITY TO MAINTAINPLURIPOTENCY IN EMBRYONIC STEM CELLS. IRANIAN RED CRESCENT MEDICAL JOURNAL (IRCMJ)[Internet]. 2018;20(8):0-0. Available from: https://sid.ir/paper/293794/en

IEEE: Copy

BEHROUZ TAHERI, MOHAMMAD HOSSEIN MODARRESSI, MAHMOUD HASHEMITABAR, MOHAMMAD MIRYOUNESI, and ELAHE MOTEVASELI, “PROKARYOTIC EXPRESSION AND PURIFICATION OF RECOMBINANT HUMANLEUKEMIA INHIBITORY FACTOR; ANALYSIS OF THE ABILITY TO MAINTAINPLURIPOTENCY IN EMBRYONIC STEM CELLS,” IRANIAN RED CRESCENT MEDICAL JOURNAL (IRCMJ), vol. 20, no. 8, pp. 0–0, 2018, [Online]. Available: https://sid.ir/paper/293794/en

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