Efficacy of Two Delivery Routes for Transplanting Human Neural Progenitor Cells (NPCs) into the Spastic Han–Wistar Rat, a Model of Ataxia

Abstract

An emerging avenue for recalcitrant neurodegenerative disease treatment is neural progenitor cell (NPC) transplantation. In this study, we investigated the effectiveness of two different delivery routes of human-derived NPC inoculation: injection into the common carotid artery or unilateral stereotactic implantation into the degenerating cerebellum and hippocampus of spastic Han-Wistar (sHW) rats, a model of ataxia. At 30 days of age, sHW mutants were implanted with osmotic pumps preloaded with cyclosporine. Ten days after pump implantation, the animals were given either 3,000,000 live human-derived NPCs (hNPCs; n = 12) or 3,000,000 dead NPCs (dNPCs; n = 12) injected into the common carotid artery, or were given two unilateral implantations of 500,000 hNPCs into the cerebellum and 500,000 hNPCs into the hippocampus of each sHW rat (n = 12) or 500,000 dNPCs by unilateral implantation into the cerebellum and hippocampus (n = 12). We also compared treated sHW rats to untreated sHW rats: normal rats (n = 12) and sibling sHW rats (n = 12). Motor activity and animal weights were monitored every 5 days to ascertain effectiveness of the two types of delivery methods compared to the untreated mutant and normal animals. Mutant rats with hNPC implantations, but not dNPC or carotid artery injections, showed significant deceleration of motor deterioration (p < 0.05). These mutants with hNPC implantations also retained weight longer than dNPC mutants did (p < 0.05). At the end of the experiment, animals were sacrificed for histological evaluation. Using fluorescent markers (Qtracker) incorporated into the hNPC prior to implantation and human nuclear immunostaining, we observed few hNPCs in the brains of carotid artery-injected mutants. However, significant numbers of surviving hNPCs were seen using these techniques in mutant cerebellums and hippocampi implanted with hNPC. Our results show that direct implantation of hNPCs reduced ataxic symptoms in the sHW rat, demonstrating that stereotactic route of stem cell delivery correlates to improved clinical outcomes.

Transplantation of Human Neural Progenitor Cells Reveals Structural and Functional Improvements in the Spastic Han-Wistar Rat Model of Ataxia

“…The other treatments of HEK and MED, in both cerebellum and hippocampus, all displayed similar level of decreased motor activity that was typical of untreated sHW rats. 15 …”

“…hNPCs were obtained according to National Institutes of Health (NIH) Ethical Guidelines and have been characterized by a previous study. 15 hNPCs were grown in culture medium consisting of animal-derived component-free (ADCF) minimum essential medium/Earle's balanced salt solution (MEM/EBSS) basal medium, supplemented with epidermal growth factor (Peprotech, Rocky Hill, NJ, USA), basic fibroblast growth factor (bFGF), insulin-like growth factor (IGF), transforming growth factor α (TGF-α), leukemia inhibiting factor (LIF) (Millipore, Temecula, CA, USA), N2 (Invitrogen, Carlsbad, CA, USA), and Gem 21 (Gemini Bioscience, Sacramento, CA, USA). The dNPCs were used to control for any paracrine effects of progenitor cell inoculations.…”

“…We also compared the effectiveness of bilateral implantations of hNPCs in the sHW rats with various controls, including dead neural progenitor cells (dNPC), a line of human embryonic kidney (HEK) cells, and human cell growth media (MED). In contrast to our previous methods study, 15 which compared intra-arterial injections with direct unilateral injections into both brain regions (cerebellum and hippocampus) simultaneously, our present study examined bilateral injections into the cerebellum or hippocampus separately. This allowed us to test the effectiveness of implanted NPCs separately in the sHW rat cerebellum and hippocampus.…”

“…13 Symptoms manifested in this animal model of ataxia are analogous to those seen in human patients, including forelimb tremors, hind-leg rigidity, gait abnormality, motor incoordination, muscle wasting, and a shortened life span (about 65 d). 14 For our first study, 15 we utilized a line of human neural progenitor cells (hNPCs), developed by Celavie Biosciences LLC (Oxnard, CA, USA) and were transplanted into the cerebellum of 40-d-old sHW rats. This recent study demonstrated that animals receiving hNPCs injections showed significant improvements in weight gain and motor activity compared to injection of dead progenitor cell controls, demonstrating the potential of these hNPCs to alleviate some symptoms caused by the sHW ataxia.…”

“…This recent study demonstrated that animals receiving hNPCs injections showed significant improvements in weight gain and motor activity compared to injection of dead progenitor cell controls, demonstrating the potential of these hNPCs to alleviate some symptoms caused by the sHW ataxia. 15 …”