Young glial progenitor cells competitively replace aged and diseased human glia in the adult chimeric mouse brain
were cryoanesthetized, secured in a custom baked clay stage and injected bilaterally with 100,000 HD glia into the presumptive striatum within 48 h of birth. Cells were delivered using a 10 μl syringe with pulled glass pipettes at a depth of 1.2–1.4 mm. The pups were then returned to their mother until weaned.
To evaluate the effects of cell age as a determinant of competitive dominance between human glia, newbornmice were injected following the same perinatal transplant protocol described above, but instead we delivered glia derived from WT-mCherry to generate human–mouse chimeras harboring WT human glia . At 40 weeks of age, WT chimeras were then injected following the same adult transplant described above, but instead, we delivered isogenic WT-EGFP glia.
Identification and phenotyping of human cells were accomplished by immunostaining for their respective fluorescent reporter, together with a phenotypic marker, including Olig2 , GFAP or Ki67 . Genetically expressed fluorescent reporters were used as markers for human cells, as their expression remained stable throughout the animal’s life .
To map the distribution and proportion of mitotically active cells within each human donor cell population, human cells expressing Ki67-immunoreactivity were mapped in every third section of the 15 equidistant sections used to perform the 3D reconstructions. Ki67 quantification was thus done every 480 µm.), the volumes of each mapped striatal section were calculated by multiplying the section thickness by the section area.
To quantify the spatial–temporal dynamics of competing human glia, we developed a program to calculate the volumetric distribution of each cell population as a function of distance to the WT glia delivery site in 3D-reconstructed datasets (Figs.
México Últimas Noticias, México Titulares
Similar News:También puedes leer noticias similares a ésta que hemos recopilado de otras fuentes de noticias.
A way forward for diagnosis of patients with extremely rare genetic mutations - Nature BiotechnologyNature Biotechnology - A way forward for diagnosis of patients with extremely rare genetic mutations
Leer más »
Van Nuys crash: 21-year-old woman killed, 4 others hurt including 11-year-old girlA 21-year-old woman was killed and several other people were hurt, including an 11-year-old girl, in a two-car crash in Van Nuys.
Leer más »
12-year-old pregnant, 18-year-old Muncie man arrested for molestationThe 12-year-old girl told police she was 6-8 weeks pregnant after she contacted police about an incident that occurred at a friend’s home
Leer más »