Two-step grafting significantly enhances the survival of foetal dopaminergic transplants and induces graft-derived vascularisation in a 6-OHDA model of Parkinson's disease.
Following transplantation of foetal primary dopamine (DA)-rich tissue for neurorestaurative treatment of Parkinson's disease (PD), only 5-10% of the functionally relevant DAergic cells survive both in experimental models and in clinical studies. The current work tested how a two-step grafting protocol could have a positive impact on graft survival. DAergic tissue is divided in two portions and grafted in two separate sessions into the same target area within a defined time interval. We hypothesized that the first graft creates a "DAergic" microenvironment or "nest" similar to the perinatal substantia nigra that stimulates and protects the second graft. 6-OHDA-lesioned rats were sequentially transplanted with wild-type (GFP-, first graft) and transgenic (GFP+, second graft) DAergic cells in time interims of 2, 5 or 9days. Each group was further divided into two sub-groups receiving either 200k (low cell number groups: 2dL, 5dL, 9dL) or 400k cells (high cell number groups: 2dH, 5dH, 9dH) as first graft. During the second transplantation, all groups received the same amount of 200k GFP+ cells. Controls received either low or high cell numbers in one single session (standard protocol). Drug-induced rotations, at 2 and 6weeks after grafting, showed significant improvement compared to the baseline lesion levels without significant differences between the groups. Rats were sacrificed 8weeks after transplantation for post-mortem histological assessment. Both two-step groups with the time interval of 2days (2dL and 2dH) showed a significantly higher survival of DAergic cells compared to their respective standard control group (2dL, +137%; 2dH, +47%). Interposing longer intervals of 5 or 9days resulted in the loss of statistical significance, neutralising the beneficial two-step grafting effect. Furthermore, the transplants in the 2dL and 2dH groups had higher graft volume and DA-fibre-density values compared to all other two-step groups. They also showed intense growth of GFP+ vessels - completely absent in control grafts - in regions where the two grafts overlap, indicating second-graft derived angiogenesis. In summary, the study shows that two-step grafting with a 2days time interval significantly increases DAergic cell survival compared to the standard protocol. Furthermore, our results demonstrate, for the first time, a donor-derived neoangiogenesis, leading to a new understanding of graft survival and development in the field of cell-replacement therapies for neurodegenerative diseases.
Laboratory of Stereotaxy and Interventional Neurosciences, Department of Stereotactic Neurosurgery, General Neurosurgery, University of Freiburg Medical Center, Freiburg, Germany; Department of Neurology, University Hospital Zurich, Zurich, Switzerland.
Fabian
F
Büchele
Laboratory of Stereotaxy and Interventional Neurosciences, Department of Stereotactic Neurosurgery, General Neurosurgery, University of Freiburg Medical Center, Freiburg, Germany.
Máté
M
Döbrössy
Laboratory of Stereotaxy and Interventional Neurosciences, Department of Stereotactic Neurosurgery, General Neurosurgery, University of Freiburg Medical Center, Freiburg, Germany; Department of Surgery, University Hospital Regensburg, Regensburg, Germany.
Christina
C
Hackl
Laboratory of Stereotaxy and Interventional Neurosciences, Department of Stereotactic Neurosurgery, General Neurosurgery, University of Freiburg Medical Center, Freiburg, Germany; Department of Neurosurgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Wei
W
Jiang
Laboratory of Stereotaxy and Interventional Neurosciences, Department of Stereotactic Neurosurgery, General Neurosurgery, University of Freiburg Medical Center, Freiburg, Germany; Federal Institute for Drugs and Medical Devices, Cellular and Systemic Neurophysiology, Bonn, Germany. Electronic address: anna.papazoglou@bfarm-research.de.
Anna
A
Papazoglou
Stereotactical Neurosurgery, University Hospital Clinics, Erlangen, Germany.
Guido
G
Nikkhah
D018663:Adrenergic Agents / Q000633:toxicity; D000818:Animals; D001058:Apomorphine / Q000494:pharmacology; D004195:Disease Models, Animal; D004298:Dopamine / Q000378:metabolism*; D059290:Dopaminergic Neurons / Q000502:physiology* / Q000637:transplantation*; D004622:Embryo, Mammalian; D005260:Female; D016332:Fetal Tissue Transplantation / Q000379:methods*; D006085:Graft Survival / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008474:Medial Forebrain Bundle / Q000293:injuries; D009412:Nerve Fibers / Q000473:pathology; D016627:Oxidopamine / Q000633:toxicity; D010300:Parkinson Disease / Q000209:etiology / Q000601:surgery*; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D013997:Time Factors
D018663:Adrenergic Agents; D049452:Green Fluorescent Proteins; D016627:Oxidopamine; D001058:Apomorphine; D004298:Dopamine
Cell survival; Cell transplantation; Graft-derived angiogenesis; Parkinson’s disease; Two-step grafting