|
Abstract Oocyte-specific linker histone(H1foo), the member of histone families, is located specificly in mammalian oocytes and primary embryos. There exists fast replacement between H1foo and its counterpart during the process of fertilization and nuclear transfer of mouse (Mus musculus) , cattle(Bos taurus) and porcine(Sus scrofa), which do help to open the chromosome in donor nuclear and achieve complete reprogramming. Induced pluripotent stem cell(iPSC) technology is another reprogramming strategy different from somatic nuclear transfer. Many reports showed that an open chromosomal structure facilitates somatic reprogramming and increases the induction efficiency of iPSCs. Therefore, in order to investigate whether H1foo has an effect on the induction efficiency of iPSCs, a transient expression of H1foo was conducted in the induction of porcine iPSCs. Two consecutive infections were conducted to the porcine fetal fibroblasts (PFF) by adding doxycycline(DOX)-inducible Lentiviral vectors expressing octamer-binding transcription factor-4(OCT4), SRY-related high-mobility-group(HMG)-box protein-2(SOX2), Kruppel-like factor-4(Klf4) and cellular homologue of avian myelocytomatosis virus oncogene(c-MYC). Vector pVenus-H1foo was electro-transferred into the OSKM-PFF and the cells transferred pVenus were acted as negative control. After several days, the obtained colonies were identified by IF and AP staining as iPSCs. By counting the number of iPSCs colonies, the results showed that the number of AP-positive colonies obtained in PFF with pVenus-H1foo or pVenus was almost the same. It was the first exploration about the effect of H1foo on induction of iPSCs which turned out there was no evident effect. The results above make basis data for the researches on mechnisms of reprogramming including somatic cell nuclear transfer and iPSCs.
|
|
Received: 03 June 2014
|
|
|
|
|
|
