Molecular Biology Of The Cell 6th Edition by Bruce Alberts, Alexander Johnson, Julian Lewis, David Morgan, Martin Raff, Keith Roberts, Peter Walter

تمرين 6

One of the earliest assays for hematopoietic stem cells made use of their ability to form colonies in the spleens of heavily irradiated mice. By varying the amounts of transplanted bone marrow cells, investigators showed that the number of spleen colonies varied linearly with dose and that the curve passed through the origin, sug- gesting that single cells were capable of forming individ- ual colonies. However, because colony formation was rare relative to the numbers of transplanted cells, it was possi- ble that undispersed clumps of two or more cells were the actual initiators. A classic paper resolved this issue by exploiting rare, cytologically visible genome rearrangements gen- erated by irradiation. Recipient mice were first irradiated to deplete bone marrow cells, and then they were irradi- ated a second time after transplantation to generate rare genome rearrangements in the transplanted cell popula- tion. Spleen colonies were then screened to find ones that carried genome rearrangements. How do you suppose this experiment distinguishes between colonization by single cells versus cellular aggregates?