In Vitro Processing Of Future Grafts

As mentioned earlier, the first limiting factor in the success of fetal grafting is the availability of suitable tissues. Banking of cryopreserved tissues is still employed and considered to be feasible by several authors. Most investigators now agree that, ideally, cryopreservation could be avoided, and optimized in vitro processing of fresh tissues for long-term survival and growth should become the standard. The University of Pittsburgh stroke transplantation trial was the first to use a cryopreserved cell line in patients. Another approach is to maintain fetal cells as aggregates in long-term suspension cultures that may even allow their expansion. However, this approach may not be feasible for processed neuronal cell lines.

There are benefits to in vitro processing ofbrain cells before implantation. First, processing may select for a viable, better characterized neuronal donor population. Second, it can help identify the donor glial— neuronal interactions that are critical in vivo, potentially protecting the graft from detrimental host reactions. Third, it can offer a setting for trophic and genetic manipulations (discussed later) that will enhance in vivo survival and integration. Finally, neuronal apoptosis in fetal grafts is well-described and appears to be significant during the first 10-15 days postimplantation. The mechanisms of neuronal death are predominantly mediated by caspases, and inhibitors of this process may have important benefits in improving the viability of the grafts.

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