Journal: PLoS ONE

Article Title: An AAV Vector-Mediated Gene Delivery Approach Facilitates Reconstitution of Functional Human CD8 + T Cells in Mice

doi: 10.1371/journal.pone.0088205

Figure Lengend Snippet: Lung, liver, spleen, kidney and bone marrow, were collected 6 and 20 weeks after NSG mice were given recombinant AAV9 vectors (5×10 9 GC of AAV9-GM-CSF, AAV9-IL-3, and AAV9-IL-15 by i.v., or 1×10 11 GC of AAV9-A2 by i.v. and intrathoracically), and DNA was isolated from the respective organs. (A) The number of AAV9 vector GC was determined by using a set of primers specific to AAV9 vector. (B) The number of GC specific for the transgenes - HLA-A2, hGM-CSF, hIL-3 and hIL-15 - was determined by using a set of primers to the respective transgene. There was a low number (10–10 3 GC; depending on the transgene) of GC detected in naïve NSG mice, as a non-specific background, and, therefore, this background GC number was subtracted from the GC number in experimental groups.

Article Snippet: Human GM-CSF, IL-3, IL-4, IL-7, and IL-15 cDNA were purchased from OriGene.

Techniques: Recombinant, Isolation, Plasmid Preparation

Journal: PLoS ONE

Article Title: An AAV Vector-Mediated Gene Delivery Approach Facilitates Reconstitution of Functional Human CD8 + T Cells in Mice

doi: 10.1371/journal.pone.0088205

Figure Lengend Snippet: (A) Schematic representation of the strategic methodology for engrafting human CD34 + cells in AAV9-transduced NSG mice. NSG mice were inoculated with AAV9-hucytokines or AAV9-A2 and 2 weeks later, mice were irradiated to myeloablate mouse immune cells. The next day, mice were transplanted i.v. with 1×10 5 human CD34 + cells previously isolated from human fetal liver. (B) The level of human CD45 + cell reconstitution in the blood was determined using flow cytometric analyses 10 weeks after engrafting human CD34 + cells into NSG mice transduced with AAV9 encoding human IL-3, IL-4, IL-7, IL-15, or GM-CSF. (C) The level of human CD45 + cell reconstitution in the blood was determined using flow cytometric analyses 10 weeks after engrafting human CD34 + cells into NSG mice transduced with individual or combination of AAV9 encoding selected human cytokines, IL-3 (5×10 9 GC), IL-15 (5×10 9 GC), and GM-CSF (1×10 9 GC). (D) The level of human CD45 + cell reconstitution in the blood was determined using flow cytometric analyses 10 weeks after engrafting human CD34 + cells into NSG mice transduced with AAV9-A2.

Article Snippet: Human GM-CSF, IL-3, IL-4, IL-7, and IL-15 cDNA were purchased from OriGene.

Techniques: Irradiation, Isolation, Transduction

Journal: MethodsX

Article Title: Automating hESC differentiation with 3D printing and legacy liquid handling solutions

doi: 10.1016/j.mex.2016.10.005

Figure Lengend Snippet: Comprehensive list of catalog numbers.

Article Snippet: IL-3 , Sino Biological , 11858-HNAE , Add to “mononuclear cell media”.

Techniques: Software