you have isolated C57BL6 embryonic stem cells and have tried anew culture condition to keep them pluripotent. To test the successof your protocol, you are injecting cells into a C57BL6 blastocyst. Once the pups are born, you notice that your experiment failed.why?

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Keith LeannonLv2

8 Mar 2019

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The second experiment is described in the following abstract:

Successful reprogramming of differentiated human somatic cells into a pluripotent state would allow creation of patient- and disease-specific stem cells. We previously reported generation of induced pluripotent stem (iPS) cells, capable of germline transmission, from mouse somatic cells by transduction of four defined transcription factors. Here, we demonstrate the generation of iPS cells from adult human dermal fibroblasts with the same four factors: Oct3/4, Sox2, Klf4, and c-Myc. Human iPS cells were similar to human embryonic stem (ES) cells in morphology, proliferation, surface antigens, gene expression, epigenetic status of pluripotent cell-specific genes, and telomerase activity. Furthermore, these cells could differentiate into cell types of the three germ layers in vitro and in teratomas. These findings demonstrate that iPS cells can be generated from adult human fibroblasts.

What does this tell you about the nature of the changes that take place during the process of differentiation? How would you think that the reactivation of these genes might act?

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In the production of iPSCs, proteins that regulate transcription are injected into differentiated cells to revert them back to stem cells. However, the efficiency of de-differentiation (ie: returning to a stem-cell like state) is typically less than 1%. Give 4 reasons why injecting the transcription factor proteins into the cell may not be enough to lead to change the cell state. In other words, why is the presence of the protein alone insufficient? Then, choose one of your 4 reasons and give a solution that would solve that problem.