Chicken leukemia inhibitory factor maintains chicken embryonic stem cells in the undifferentiated state.
Mouse embryonic stem (ES) cells can be maintained in an undifferentiated state in the presence of leukemia inhibitory factor (LIF), a member of the interleukin-6 cytokine family. In other mammals, this is not possible with LIF alone. Chicken ES-like cells (blastodermal cells) have only been cultured with mouse LIF because chicken LIF was not available. However the culture system is imperfect and chicken ES-like cells equivalent to mouse ES cells were not observed. In the present study, we cloned the cDNA-encoding chicken LIF using mRNA subtraction and RACE methodology. The chicken LIF cDNA encodes a protein with approximately 40% sequence identity to mouse LIF. It has 211 amino acids including a putative N-terminal signal peptide of 24 residues. Chicken blastodermal cells were cultured in the presence of bacterially expressed chicken LIF or mouse LIF. The expression of alkaline phosphatase and embryonal carcinoma cell monoclonal antibody-1 and stage-specific embryonic antigen-1 and the activation of STAT3 were examined, all of which are indices of the undifferentiated state. Exposure in the blastodermal cells to recombinant chicken LIF but not to mouse LIF maintained the expression of these various markers. After 9 days of incubation, the blastodermal cells formed cystic embryoid bodies in the presence of mouse LIF but not in the presence of recombinant chicken LIF. We conclude that chicken LIF is able to maintain chicken ES cell cultures in the undifferentiated state.
Horiuchi
Hiroyuki
H
Department of Molecular and Applied Biosciences, Graduate School of Biosphere Science, Laboratory of Immunobiology, Hiroshima University, Higashi-Hiroshima 739-8528, Japan. hhori10@Hiroshima-u.ac.jp
Tategaki
Airo
A
Yamashita
Yusuke
Y
Hisamatsu
Hikaru
H
Ogawa
Mari
M
Noguchi
Takashi
T
Aosasa
Masayoshi
M
Kawashima
Tsuyoshi
T
Akita
Sachiko
S
Nishimichi
Norihisa
N
Mitsui
Naoko
N
Furusawa
Shuichi
S
Matsuda
Haruo
H
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism; D015153:Blotting, Western; D018236:Carcinoma, Embryonal / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian / Q000166:cytology*; D004625:Embryo, Nonmammalian; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism / Q000502:physiology*; D053605:Leukemia Inhibitory Factor; D020935:MAP Kinase Signaling System; D051379:Mice; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010766:Phosphorylation; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013234:Stem Cells / Q000166:cytology*; D013997:Time Factors
D018076:DNA, Complementary; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D021382:Protein Sorting Signals; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000469:Alkaline Phosphatase