Human embryonic dopamine neurons xenografted to the rat: effects of cryopreservation and varying regional source of donor cells on transplant survival, morphology and function

Frodl, Eva M.; Duan, Wenjun; Sauer, H.; Kupsch, Andreas; Brundin, Patrik

doi:10.1016/0006-8993(94)91328-5

Cited by 79 publications

(29 citation statements)

References 39 publications

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“…The physiological function of the sorted cell population was then analyzed by transplanting the cells into a rat model of Parkinson's disease. Considering both the proportion of DA neurons in the sorted cells and the cell count in the graft, the survival rate of the DA neurons after intrastriatal transplantation can be estimated to be 6-10%, which is similar to those found in xenograft experiments (28,29). Furthermore, the surviving DA neurons extensively innervated the host striatum.…”

Section: Discussionsupporting

confidence: 63%

Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Sawamoto

Nakao

Kobayashi

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

219

209

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To visualize and isolate live dopamine (DA)-producing neurons in the embryonic ventral mesencephalon, we generated transgenic mice expressing green fluorescent protein (GFP) under the control of the rat tyrosine hydroxylase gene promoter. In the transgenic mice, GFP expression was observed in the developing DA neurons containing tyrosine hydroxylase. The outgrowth and cue-dependent guidance of GFP-labeled axons was monitored in vitro with brain culture systems. To isolate DA neurons expressing GFP from brain tissue, cells with GFP fluorescence were sorted by fluorescence-activated cell sorting. More than 60% of the sorted GFP ؉ cells were positive for tyrosine hydroxylase, confirming that the population had been successfully enriched with DA neurons. The sorted GFP ؉ cells were transplanted into a rat model of Parkinson's disease. Some of these cells survived and innervated the host striatum, resulting in a recovery from Parkinsonian behavioral defects. This strategy for isolating an enriched population of DA neurons should be useful for cellular and molecular studies of these neurons and for clinical applications in the treatment of Parkinson's disease.

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Section: Discussionsupporting

confidence: 63%

Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Sawamoto

Nakao

Kobayashi

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

219

209

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“…Female Sprague-Dawley rats were subjected to unilateral 6-hydroxydopamine lesions of the ascending mesostriatal dopamine pathway as described (5). The effect of the 6-hydroxydopamine lesion was assessed by monitoring amphetamine (2.5 mg/kg, i.p.…”

Section: Methodsmentioning

confidence: 99%

“…However, the results obtained so far indicate that the symptomatic relief is far from complete and it has been suggested that improved functional effects would be attainable if the grafts contained more surviving dopamine neurons and innervated a larger volume of the denervated parkinsonian striatum (1). Experiments with transplants of rat and human mesencephalic dopamine neurons placed in the rat striatum have shown that on the order of 5-20% of the dopamine neurons that are dissected from the embryos survive the stereotaxic implantation procedure when a protocol similar to that employed clinically is used (2)(3)(4)(5). The underlying reason for the relatively poor survival rate of grafted dopamine neurons is not known.…”

mentioning

confidence: 99%

“…The cell viability of the cell suspensions just prior to grafting was 92-94% in all groups. The cell concentrations in the control, U-74389G-treated, and U-83836E-treated, groups were (5).…”

mentioning

confidence: 99%

“…Cell suspensions for both in vitro viability testing and intracerebral transplantation were prepared from ventral mesencephalic tissue obtained from embryonic day 14 rat embryos (Sprague-Dawley; crown-to-rump length, 12-13 mm) as described (5). In the transplantation experiment, two pregnant rats were injected (10 mg/kg, i.p.)…”

mentioning

confidence: 99%

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Lazaroids improve the survival of grafted rat embryonic dopamine neurons.

Nakao

Frodl

Duan³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

208

100

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In rodent models of Parkinson disease in which transplants of dissociated rodent and human embryonic mesencephalic tissue, rich in dopamine neurons, have been studied, only 5-20% of the dopamine neurons survive the implantation procedure. We have investigated the effects of inhibiting free radical generation with two lazaroids, U-74389G and U-83836E, on the survival of embryonic rat dopamine neurons. U-74389G is a 21-aminosteroid, and U-83836E combines the piperazinyl pyrimidine portion of 21-aminosteroids with the antioxidant ring of a-tocopherol. In an initial study, we found that the lazaroids markedly prolonged the period after tissue dissociation that an embryonic mesencephalic cell suspension exhibits high cell viability in vitro, as assessed by using a dye exclusion method. In a second series of experiments, addition of lazaroids to dissociated mesencephalic graft tissue increased the yield of surviving rat dopamine neurons 2.6-fold after implantation in the dopamine-denervated rat striatum. The improved survival correlated with an earlier onset of graft-induced functional effects in the amphetamineinduced rotation test. Thus, inhibition of free radical generation can significantly increase the yield of grafted embryonic dopamine neurons. Addition of lazaroids to the graft preparation is a relatively simple modification of the transplantation protocol and could readily be applied in a clinical setting. Moreover, since iron-dependent lipid peroxidation has been suggested to play a role in the death of nigral dopamine neurons in Parkinson disease and lazaroids are particularly potent inhibitors of such processes, the findings may have implications for the pathogenesis of this disease.Several studies indicate that grafts of human embryonic dopamine neurons can survive and reduce motor symptoms after transplantation to the brains of patients with Parkinson disease (1). However, the results obtained so far indicate that the symptomatic relief is far from complete and it has been suggested that improved functional effects would be attainable if the grafts contained more surviving dopamine neurons and innervated a larger volume of the denervated parkinsonian striatum (1). Experiments with transplants of rat and human mesencephalic dopamine neurons placed in the rat striatum have shown that on the order of 5-20% of the dopamine neurons that are dissected from the embryos survive the stereotaxic implantation procedure when a protocol similar to that employed clinically is used (2-5). The underlying reason for the relatively poor survival rate of grafted dopamine neurons is not known. However, we have hypothesized (6) that the major loss of dopamine neurons occurs either during dissection and mechanical dissociation of the graft tissue or soon after implantation into the adult brain environment.

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Comparison of fresh and cryopreserved porcine ventral mesencephalon cells transplanted in A rat model of Parkinson's disease

Jacoby¹,

Lindberg²,

Ratliff³

et al. 2002

J of Neuroscience Research

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To evaluate whether cryopreservation of porcine ventral mesencephalon cells influences graft survival and function in vivo, we have transplanted either freshly prepared or cryopreserved cells into the striatum of 6-hydroxydopamine-lesioned rats. A single cell suspension of porcine ventral mesencephalon cells from the same isolation either was stored at 4 degrees C and transplanted the next day or was cryopreserved for 4 weeks in liquid nitrogen vapor. The cryopreserved cells were then rapidly thawed, rinsed, and transplanted in the same manner as the fresh cells, with the same dose of viable cells. All animals received daily injections of cyclosporin A to prevent xenograft rejection. To monitor graft function, amphetamine-induced rotation was measured every 3 weeks between 6 and 15 weeks posttransplantation. After sacrifice at 15 weeks posttransplantation, histological methods were used to compare fresh cell and cryopreserved cell transplants with respect to graft survival, differentiation and integration, and host immune response. Cryopreserved cells were found to be either equivalent or in some cases superior to fresh cells with respect to rotational correction, graft survival, graft volume, numbers of graft-derived dopaminergic neurons, and host immune responses. In conclusion, the results indicate that it is feasible to cryopreserve porcine ventral mesencephalon cells for long-term storage of cells prior to transplantation in an animal model of Parkinson's disease.

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Human embryonic dopamine neurons xenografted to the rat: effects of cryopreservation and varying regional source of donor cells on transplant survival, morphology and function

Cited by 79 publications

References 39 publications

Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Visualization, direct isolation, and transplantation of midbrain dopaminergic neurons

Lazaroids improve the survival of grafted rat embryonic dopamine neurons.

Comparison of fresh and cryopreserved porcine ventral mesencephalon cells transplanted in A rat model of Parkinson's disease

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