2000;97:14720C14725. as described previously [4C7]. Prior to transplantation, rats (= 71) were randomized across cohorts, and comparative behavioral baselines were confirmed for each cohort using pretransplant Basso, Beattie, and Bresnahan (BBB) locomotor scores 7 or 55 DPI (observe also Behavioral Assessments). Animals with abnormal scores (>2 SDs outside S130 the cohort mean), unilateral bruising, or abnormal pressure/displacement curves after contusion injury, or in which a vertebral T9 laminectomy could not be confirmed at the time of cell transplantation, were excluded from the study. An S130 additional eight animals were lost because of anesthesia/surgery-related complications. After these exclusions behavioral and histological assessments were completed in = 47 animals. All animal care, behavioral assessments, and histological processing/analysis were performed by observers blinded to the experimental cohorts. hCNS-SCns engraftment was confirmed in all animals, and a subset of spinal cords from animals in all cohorts was randomly selected for stereological analysis. SC121 immunostaining revealed that 2 of 9 rats from your 9 DPI hCNS-SCns cohort and 4 of 11 rats from your 60 DPI hCNS-SCns cohort showed very poor or no engraftment; these rats were excluded from further sensory behavioral and stereological analysis, and from statistical analysis other than reporting of the percentage of engrafted animals. Final cohort figures (= 10; 9 DPI vehicle, = 12; 60 DPI hCNS-SCns, = 7; 60 DPI vehicle, = 12. Final cohort figures for histology/stereology or evaluation of locomotor function at 14 weeks post-transplantation (WPT) were as follows: 9 DPI hCNS-SCns, = 7; 9 DPI vehicle, = 8; 60 DPI hCNS-SCns, = 7; 60 DPI vehicle, = 12 (supplemental online Table 1). Behavioral Assessments Mechanical allodynia assessment using von Frey screening [26] and thermal hyperalgesia assessment using Hargreaves screening [25] were conducted prior to injury (baseline), and at 2, 7, 11, and 14 WPT as explained in [7]. CatWalk video of three individual runs per animal was recorded at 14 WPT and analyzed using CatWalk software version 6.13 for Windows by individuals blinded to experimental groups [7]. Hind limb base S130 of support steps are shown relative to baseline obtained in uninjured ATN rats assessed at 7 BII weeks of age. BBB open-field screening was performed as published by Basso et al. [13]; however, we found assessment of coordination around the BBB in the ATN rat strain to be flawed. Specifically, the number of passes in which locomotion was carried out at a consistent velocity for an assessable distance was too low to achieve an acceptable degree of accuracy, regardless of the amount of habituation to the task the animals received or manipulation of task parameters. Pretransplantation BBB scores were not critically affected because they ranged below the affected portion of the BBB level, particularly in the case of the 9 DPI cohort. However, by several weeks post-transplant, a majority of the animals were performing within the range of the BBB rating level where accurate assessment of coordination was crucial. To address this issue, we used the 14 WPT CatWalk data to establish a coordination score for the 14 WPT BBB data, as previously explained by Hamers et al. [31]. Using this method,.