Vp10m200st

CBF was monitored before, during and after surgery using a laser-Doppler flowmeter (moorVMS-LDF1, Moor Instruments, Devon, UK). A ‘master’ probe (#VP10M200ST, Moor Instruments, Devon, UK), attached at one end to the laser-Doppler, was attached at the alternate end to a ‘slave’ probe (#P10d, Moor Instruments, Devon, UK). The slave probe was then fed through a silicon electrode holder (#PHDO, Moor Instruments, Devon, UK). Four drops of Loctite Gel (#10762, Loctite) were applied to the silicon holder, just surrounding, but not touching, the slave probe. Optical matching gel (#PMF, Moor Instruments, Devon, UK) was applied to the tip of the slave probe. The silicone holder/slave probe was adhered perpendicular to the right temporal skull at Antero-Posterior (AP) ~-1.0 and Medio-Lateral (ML) ~3.0 from bregma, to measure blood flow in the territory supplied by the right MCA [24 (link)] (Fig 1A). Once Loctite Gel firmed the mice, with the laser-Doppler now attached, were carefully released from the stereotaxic apparatus and gently placed supine upon the surgical table, with an area cut from the styrofoam platform to allow room for the attached laser-Doppler probe.

This study was conducted on adult male IL-1Ra transgenic (Tg) mice (hemizygous mice carrying the transgene encoding secreted IL-1Ra mRNA under the control of its endogenous promoter) and LM controls [11 (link),28 (link)]. Transient (t)MCAo was performed under 1–2% isoflurane anesthesia. Mice received 1 mL (s.c.) 0.9% saline before the transient intraluminal filament technique (45 min) [29 (link)]. Blood flow and body temperature were monitored using an optical fiber probe (T6a and VP10 M200st, Moor Instruments, UK) connected to a laser Doppler flowmeter (Moor Instruments, UK). Postsurgery, the mice received 0.1 mL (s.c.) 5% glucose and 0.1 mL Temgesic (buprenorphium 0.3 mg/mL; pharmaceuticals, USA). Temgesic was given at 8 h intervals for the first 24 h poststroke [11 (link)].
BM harvesting, characterization, and treatment have previously been detailed [8 (link),11 (link)]. Approximately 1 × 10⁷ BM cells were injected into the tail vein of recipient mice 30 min after tMCAo [11 (link)]. In this study, we include samples from LM mice subjected to tMCAo (LM) (n = 10), LM mice subjected to tMCAo, followed by LM BM treatment (LM–LM) (n = 12), or IL-1Ra BM treatment (Tg–LM mice) (n = 12), and unlesioned controls (Ctl) (n = 11) [11 (link)].

All studies were conducted using young age-matched male mice. Focal cerebral ischemia was induced by either permanent (p) or transient (t) middle cerebral artery occlusion (MCAo). pMCAo permanent occlusion of the distal part of the left MCA was performed as detailed in Clausen et al. [10 (link)]. tMCAo transient occlusion (40 min) of the right MCA was performed using the intraluminal filament placement technique, as detailed in Inacio et al. [35 (link)], under 1.5 % isoflurane anesthesia. All studies were conducted as randomized, double-blinded studies, with inclusion of sham-operated mice and unlesioned controls (Ctl). Blood flow and body temperature were monitored using the optical fiber probe (VP10 M200ST, UK) and endoscopic temperature probe (T6a, UK) from Moor Instruments. The study was approved by the Danish Veterinary and Food Administration (J. no. 2011/561-1950). An overview of the different mouse groups used for assessment of the effect of IL-1Ra and IL-1Ra-producing cells on infarct volume is provided in Table S1.