Stem cell counting: Development and application

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The stem cells of the blood-forming system (hematopoietic stem cells, HSCs) are rare, comprising only 1 in every 20,000 bone marrow cells. Because they are the only cell in marrow with the capacity to sustain blood cell production throughout adult life, they play a central role in many blood cell disorders and most leukemias. They are also the principal therapeutic entity responsible for the regenerative effectiveness of marrow or cord blood transplants. There is accordingly a need to be able to enumerate them in many circumstances of practical clinical importance. Outcomes of clinical HSC transplants, for example to treat blood diseases such as leukemia, depend critically on the number of HSCs in the donor sample. Unfortunately, there presently exists no practical means of counting HSCs directly. Instead, researchers that need to enumerate them must resort to extremely cumbersome and expensive assays that involve injecting the cells into mice and then waiting a minimum of 5 months before obtaining a result. The enormous effort, time and cost it takes to enumerate HSCs are major limiting factors in blood stem cell research and clinical practice.We propose here the development of a technology for direct and rapid marker-based enumeration of HSCs within half a day, in any cellular sample regardless of biological state and without requirement for prior enrichment. Our proposal brings together a multidisciplinary team of world experts in the fields of stem cell bioengineering (Zandstra), materials and microfluidics (Kumacheva), stem cell biology (Iscove) and single-cell cytometry (Audet). The anticipated outcome will be a high-throughput microfluidics automatable system (Stem Cell Counting Chip - SCCC) which will be translated, with the help of Centre for the Commercialization of Regenerative Medicine and STEMCELL Technologies, into a device usable in any hospital for the robust enumeration of rare cell types such as HSCs. We expect the SCCC to revolutionize stem cell research and regenerative medicine by providing the tools to enumerate HSCs in donor samples or in patients, in real time.