Cell Constituents Adrenal Medulla are Detrimental Survival ... · maffin cells in transplantation experiments was to provide a paraneural source of dopamine to replenish the dopamine

Non-Chromaffin Cell Constituents of the Adrenal Medulla areDetrimental to the Survival of Grafted Adrenal Chromaffin

Cells: Studies in Rats and Non-Human PrimatesSherry B. Schueler, John D. Ortega, Jacqueline Sagen, and Jeffrey H. Kordower

Department ofAnatomy and Cell Biology, University ofIllinois School ofMedicine, andDepartment ofNeurological Sciences, Rush Presbyterian-St. Luke’s Medical Center, Chicago, Illinois 60612, USA

The initial rationale for using adrenal chro-maffin cells in transplantation experiments wasto provide a paraneural source of dopamine toreplenish the dopamine insufficiency created inanimal models of parkinsonism and idiopathicParkinson’s disease. Intraventricular transplantsof adrenal medulla (AM) survive, synthesize andsecrete eatecholamines, and reverse drug-in-duced motor asymmetries in unilateral nigrostri-atal lesioned rats. Subsequent studies in whichAM grafts were placed directly into the striatumalso induced functional recovery, albeit partially,and short-lived. It soon became increasinglyclear that ehromaffin cells survived poorly withinthe striatal parenchyma. Studies carried out innon-human primates and autopsy eases fromAM-grafted parkinsonian patients confirmed thenotion that adrenal ehromaffin cells do not sur-vive well following intrastriatal transplantation.

The use of trophic factors to improve chro-maff cell survival heralded the second era ofAM transplantation. Injections of /3. nervegrowth factor into AM graft sites augmentschromaffin cell survival, induces morphologicaldifferentiation, and increases the magnitude andduration of functional effects. Chromaffin cellsurvival can also be increased by cografting AMwith growth factor-producing C6 gliomas, astro-cytes genetically engineered to produce/3NGF,and transected peripheral nerve whose damagedSchwann cells secrete a variety of trophicmolecules including/3NGF.We have begun to utilize an alternative ap-

proach towards enhancing chromaffin cell sur-vival. Instead of using trophic molecules to en-hance graft viability we have attempted to mini-mize factors which may negatively impact uponchromaffin cell survival and induce AM grafts to

degenerate. Adrenal chromaffin cells have pre-viously been demonstrated to survive well fol-lowing implantation into the periaqueductal grayonce the chromaffin cells were isolated from non-chromaffin cells found in the AM. These datasuggest that fibroblasts, blood-borne leukocytes,and/or endothelial cells within the AM may bedetrimental to chromaffin cell viability and, atleast in part, underlie the poor survival of thesecells following intrastriatal implantation.We presently assessed the effects of isolating

bovine ehromaffin cells from the other cell typeswithin the AM upon ehromaffin cell graft sur-vival in rats and MPTP treated rhesus monkeysfollowing intrastriatal transplantation. Threegroups of immunosuppressed rats were em-ployed. Group 1 received grafts of bovine AMfollowing gland perfusion and dissection of themedulla from the adrenal cortex. Group 2 re-ceived implants of isolated adrenal chromaffincells. Chromaffin cells were isolated in vitro byplacing them in a Pereoll gradient resulting inthe segregation of dead cells, residual corticalcells, red blood cells, and viable medullary cells.Once separated, the medullary cells were differ-entially plated resulting in a 95% pure popula-tion of chromaffin cells. Group 3 received im-plants of isolated adrenal chromaffin cells whichwere reseeded with fibroblasts and endothelialcells which were allowed to proliferate in cul-ture. All rats were sacrificed 1-2 months follow-ing transplantation. Rats in Group 1 displayedviable tyrosine hydroxylase (TH) and dopamine/3 hydroxylase (D/3H)-immunoreactive (ir)transplants. However, these grafts tended to besmall, the graft-host interface was infiltratedwith macrophages, and many of the chromaffincells appeared to be in the process of degenera-

(C) FREUND PUBLISHING HOUSE LTD., LONDON. JOURNAL OFNEURALTRANSPLANTATION & PLASTICITY, Vol. 3, No. 4,1992

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tion. In contrast, rats receiving implants of iso-lated chromaffin cells displayed very large andhealthy appearing TH-ir and DflH-ir transplantswhich retained their endocrine phenotype. Ratsin this group displayed a 2- and 3-fold increase insurviving chromaffin cells one and two monthsfollowing transplantation, respectively, relativeto rats in Group 1. Isolated chromaffin cell im-plants appeared to integrate well within the hostand few macrophages were evident. The re-seeding of isolated chromaffin cells with fibrob-lasts and endothelial cells impaired chromaffincell survival. Most rats in this group failed to dis-play surviving implants and the few survivingchromaffin cells appeared to be degenerating.Interestingly, the perigraft region in rats fromthis group displayed a dense TH, but not DflH,fiber network, suggesting that signals from de-generating implants may modify the host nigros-triatal system.

To assess the ability of isolated adrenal chro-maffi cells to survive grafting in the primatebrain, these cells were implanted into the cau-date and putamen of immunosuppressed hemi-parkinsonian rhesus monkeys. One month fol-lowing transplantation, robust survival(> 1,000,000) of endocrine appearing TH- andD/3H-immunoreactive chromaffin cells was ob-served. These cells appeared to integrate wellwithin the striatum.

These data strongly suggest that non-chro-maffi cell constituents within the AM aredetrimental to the survival of chromaffin cellsfollowing intracerebral transplantation. Isolatingchromaffin cells prior to transplantation maysignificantly enhance their survival and may bean improved donor source for the treatment ofParkinson’s disease.

Supported by UPF and NS28931.

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Cell Constituents Adrenal Medulla are Detrimental Survival ... · maffin cells in transplantation experiments was to provide a paraneural source of dopamine to replenish the dopamine