Establishing lymphoblastoid cell lines from frozen blood of extremely old individuals

In population studies involving peripheral blood samples from nonagenarian and centenarian donors, the amount of biological material available for research is restricted, as only a few millilitres of blood can be obtained from extremely old donors without significantly compromising their health. Here we describe a protocol to immortalize small amounts of total frozen blood from extremely old donors (90+) using the Epstein-Barr virus, despite the low level of circulating B cells present in nonagenarian and centenarian blood samples. This methodology provides a unique way to maximize resources of biological material available for research.