Hypoimmune induced pluripotent stem cells survive long term in fully immunocompetent, allogeneic rhesus macaques
Approximately 20 μl of aggregates was collected into a 1.5-ml Eppendorf tube, washed with DPBS, fixed in 4% paraformaldehyde in DPBS for 1 h at 4 °C, washed with DPBS and stored in DPBS at 4 °C until staining . Aggregates were incubated in 100% methanol pre-cooled at −20 °C for 30 min, and then 900 μl of methanol was replaced with 900 μl of DPBS. Samples were spun down and washed twice with DPBS.
To induce diabetes, animals received daily intraperitoneal injection of streptozotocin for five consecutive days at 60 mg kgbody weight in 0.1 M citrate buffer. Glucose levels were determined by glucometer in 10 μl of blood samples collected by tail vein venipuncture after 4 h of fasting. Animals with a glucose level of more than 450 mg dlwere served as recipients for cell implant.
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