Wt c57bl6 nj mice

The mice were housed in a temperature (23 ± 1°C)- and humidity-controlled facility under a 12-h dark/light cycle, fed standard mouse chow (Prolab IsoPro RMH 3000, #3005737-220, Lab Diet) and water ad libitum. The experiments were performed on adult male mice (12–16 weeks old). Bradykinin B1 receptor knockout (B1RKO) mice were a generous gift from Dr. Michael Bader (Charité Hospital, Berlin, Germany) and originated from the backcrossing of an initially mixed genetic background (129/Sc and C57Bl/6) with C57Bl/6 mice (13 (link)). Wildtype (WT) C57Bl/6NJ mice (stock no. 005304) were purchased from the Jackson Laboratory. All animal studies were approved by the East Carolina University Animal Care and Use Committee (AUP #W254) and were performed in accordance with the National Institutes of Health (NIH) Guidelines for the Care and Use of Laboratory Animals and ARRIVE guidelines.

All experiments were carried out with proper oversight by institutional review boards for animal care. Comparisons of i.v. and i.c.v. routes of administration were completed using wild-type (WT) C57BL/6J mice (Jackson Laboratory, stock number 000664). Proof-of-concept experiments for GBA-PD were completed using homozygous Gba^D409V/D409V mutant mice (Jackson Laboratory, C57BL/6N-Gba^tm1.1Mjff/J, stock number 019106)³⁹ (link) and WT C57BL/6NJ mice (Jackson Laboratory, stock number 005304) as controls. Proof-of-concept experiments for GRN-FTD were completed using heterozygous and homozygous Grn^R493X mutant mice on the C57BL/6J background.⁴¹ (link) Donor and recipient animals for all experiments were 8–12 weeks of age. Terminal collection for all animals occurred 14–16 weeks after cell administration except for the long-term cohort for scRNA-seq experiments (12–13 months post-transplantation) and the long-term cohort for the GRN-FTD proof-of-concept study (32–36 weeks post-transplantation). At necropsy, animals first underwent whole-body transcardial perfusion with either heparinized saline or PBS followed by tissue harvesting. Samples for biochemistry analysis were flash frozen and stored at -80°C. Samples for immunohistochemistry (IHC) analysis were fixed in 4% PFA in PBS overnight at 4°C.