Labeling efficacy of superparamagnetic iron oxide nanoparticles to human neural stem cells: comparison of ferumoxides, monocrystalline iron oxide, cross-linked iron oxide (CLIO)-NH2 and tat-CLIO.

OBJECTIVE

We wanted to compare the human neural stem cells (hNSC) labeling efficacy of superparamagnetic iron oxide nanoparticles of different (SPIONs), namely, ferumoxides, Human Clia kits monocrystalline iron oxide (MION), cross-linked iron oxide (CLIO) -NH (2) and tat -CLIO ,

METHOD

The hNSCs (5 x 10 (5) HB1F3 cells / ml) were incubated for 24 hours in cell culture media containing 25 microg / ml ferumoxides, MION or CLIO-NH (2), and with or without poly-L-lysine (PLL ) and tat-CLIO. Cellular iron absorption was analyzed qualitatively by using a light microscope and measured by atomic absorption spectrophotometry. Visibility of the labeled cells was assessed by MR imaging.

RESULTS

SPIONs incorporation into hNSCs not affect cell proliferation and viability. The hNSCs labeled with tat-CLIO show the longest retention, up to 72 hours, and they contained iron of 2.15 +/- 0.3 pg / cell, 59-fold, 430-fold and six times more iron than hNSCs included labeled ferumoxides, MION or CLIO-NH (2), respectively. However, when the PLL is added, merging ferumoxides, MION or CLIO-NH (2) into hNSCs is comparable with tat-CLIO.

CONCLUSION

For MR imaging, hNSCs can be efficiently labeled with tat-CLIO alone or in combination with ferumoxides, MION, CLIO-NH (2) and transfection agent PLL.

Quantitative Proteomic Analysis of Human Airway Cilia Identify Protein Previous uncharacterized of High Abundance.

Cilia are very important to many diverse cellular processes. Although many major axonemal components have been identified and studied, how they interact to form a functional axoneme is not fully understood. To further our understanding of the composition of the human respiratory tract cilia proteins, we performed semiquantitative analysis of axonemes cilia using label-free LC / MSE, identified more than 400 proteins with high confidence. Tubulin is the Mouse Clia Kits most abundant proteins are identified, with evidence of 20 different isoforms obtained. 

Twelve different isoforms axonemal dynein heavy chain were also identified. absolute quantification nontubulin components showed greater than 75-fold range of protein abundance (RSPH9; 1850 fmol vs. CCDC103; 24 fmol), adding another level of complexity axonemal structure. Of the identified proteins, ~ 70% protein known axonemal. In addition, many previously uncharacterized proteins identified. Unexpectedly, some of these, including ERICH3, C1orf87, and CCDC181, present at relatively high abundance in cilia.