I had previously written a little about Stem cells. While researchers still don’t yet know exactly how the four factors transform the fully differentiated fibroblast cells back into pluripotency, possible explanations are pouring in.
Pluripotency (by which the stem cell may become any tissue; muscle or nerve, for example) and “self renewal” (cells should not only differentiate, some ready stock of stem cells must be there for future need) are important determinants for stem cells.
According to Shinya Yamanaka, the steps could be somewhat like this: c-Myc first confers the open chromatin state and immortality to the skin fibroblasts. But it also induces apoptosis by acting on the p53, “the guardian of the genome”. Apoptosis or cellular senescence causes the cells to die. Klf4 inhibits p53 induced apoptosis. Again, if we added only Klf4 and c-Myc we would get tumor cells (both being oncogenes). Oct4 here acts and makes ES like cells (ES= Embryonic Stem) out of what was destined to be tumor cells. Sox2 confers pluripotency and you’ve got what you wanted.
Now, we just have to hand pick the right cells from the petridish. Scientists can do it either by looking for Fbx15 expression or the expression of nanog in the treated sample. Both Fbx15 and Nanog are targets of Oct3/4 and Sox2; but Nanog is found to be more closely associated with pluripotency, as is evidenced by adult chimera formation (chimera is a monstrous fire breathing creature like dragon of ancient mythology).
There have been some important modifications. Researchers have shown that one could still get human induced pluripotent stem cells (iPSC) without the need of the c-myc oncogene. The mode of delivery of these four factors could also be undertaken by plasmids, rather than the traditional retroviral vector approach. Retroviruses (like c-Myc) could potentially induce cancer. You may like to hear this Nature Podcast where both Yamanaka and Rudolph Jaenisch give a very good summary. As a bonus, you may also appreciate another way of creating iPSC. Replace the genome in “early embryonic cells” or zygotes (fertilized eggs) during cell division. During cell division, the nuclear membrane disappears and the factors are no longer in the nucleus. They are in the cytoplasm. Dieter Egli explains that if you replaced the genome of this zygote with another (genome) while the cell was still dividing, the new genome would adapt to the new cytosolic environment and get instructions from the factors in the cytosol. It will go ‘back in time’ and become a stem cell.
Now, a bit of refreshment. Watch this awe inspiring Metallica video called 'All nightmare long'. It portrays the Tunguska event, A-bomb, Soviet Revolution, American supremacy (?) and ‘revival of organisms’. Some key phrases are:
- “like a split worm, a part of the organism can reconstitute the whole”. Check about Planarians (flat worms, picture on the left), they not only reconstitute but also become separate individuals!
- “Instead of offspring, they become skin cells, nerves and muscle”- just as we described! Seems Metallica is well informed! Do see this wonderful video in YouTube (Metallica All Nightmare Long (Official Music Video))
Okita, K., Ichisaka, T., & Yamanaka, S. (2007). Generation of germline-competent induced pluripotent stem cells Nature, 448 (7151), 313-317 DOI: 10.1038/nature05934
Developmental reprogramming after chromosome transfer into mitotic mouse zygotes, doi:10.1038/nature05879