Application of a High Efficiency Retroviral Infection Technique for the Generation of Marmoset Induced Pluripotent Stem Cells

Thomas Focht

Mentor: Dr. Peter Hornsby

The goal of Dr. Hornsby’s research is to make induced pluripotent stem cells (iPS cells) from the common marmoset (any of several small, squirrel-like, South and Central American monkeys of the genera Callithrix, Leontocebus, etc.). The discovery of that would allow him to understand the safety and effectiveness of therapies using iPS cells. He has established a way to produce marmoset iPS cells by transferring the genetic material of four transcription factors that control the transfer of genetic material. Dr. Hornsby’s investigation entailed infecting cells from connective tissues with retroviruses by centrifuging the cells with the groups of retrovirus. This infection was repeated twice in 48 hours. After infection, the cells were replated in a supplement with growth factors. The following day cells were changed to a new nutrient supplement called mTesR1 that has special contents. For 12 days the cells were treated with another nutrient supplement and an agent that speeds up reprogramming. At about 1 week, cells with an altered appearance were observed in a nutrient supplement. At about 2 weeks of being in the nutrient, there were colonies of cells believed to be iPS cells observed. The current objective is to isolate and take out those colonies for future experimentation.

Collaborators: Yuehong Wu