Isolation of Human Dermal Fibroblasts from Biopsies

Although new methods for generating iPSCs have been a hot topic for the past several years, less attention has been focused on the cells being reprogrammed. Fibroblasts have been the most common source of reprogrammable cells, probably because there are several resources that have already banked fibroblasts from both diseased and normal donors. However, there is a growing need for primary cells that are not available from a company or a somatic cell bank. When considering banking their own collections of somatic cells for reprogramming, researchers need to consider which somatic cell type would be the best for their particular application. Over the years, a number of different somatic cell types have been successfully reprogrammed into iPSCs with varying efficiency, including fibroblasts, keratinocytes from hair and skin, various cells from the blood, and urothelial cells from urine. Each cell type has different advantages and disadvantages. For example, isolating keratinocytes from the outer root sheath of hair does not require a physician’s assistance, is painless for the donor, and the cells reprogram with high efficiency. However, primary keratinocytes do not expand well nor do they recover well after cryopreservation. Blood is fairly easy to acquire but the reprogramming efficiency is low and since normal blood cells do not proliferate well in vitro , there is limited potential for banking cells for further reprogramming. In our lab, we have generated a number of iPSC lines using different types of primary cells that we isolated ourselves. We have weighed the pros and cons of each cell type and have found that human dermal fibroblasts (HDF) are the most robust cell type for reprogramming purposes with the fewest drawbacks.