Thursday, September 17, 2009

9/18 pubmed: adipose stem cell

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Cryo-Imaging of Fluorescently-Labeled Single Cells in a Mouse.
September 17, 2009 at 7:52 am

Cryo-Imaging of Fluorescently-Labeled Single Cells in a Mouse.

Proc Soc Photo Opt Instrum Eng. 2009 Jan 1;7262:72620W-72620W8

Authors: Steyer GJ, Roy D, Salvado O, Stone ME, Wilson DL

We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryo-imaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 mum) and imaging, collecting color brightfield and fluorescent block-face image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [u(T) : heart (267 +/- 47.6 mum), liver (218 +/- 27.1 mum), brain (161 +/- 27.4 mum)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns.We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron-scale, fluorescence, and bright field image data. Here we describe our image pre-processing, analysis, and visualization techniques. Processing improves axial resolution, reduces subsurface fluorescence by 97%, and enables single cell detection and counting. High quality 3D volume renderings enable us to evaluate cell distribution patterns. Applications include the myriad of biomedical experiments using fluorescent reporter gene and exogenous fluorophore labeling of cells in applications such as stem cell regenerative medicine, cancer, tissue engineering, etc.

PMID: 19756213 [PubMed - as supplied by publisher]


Comparison of adult neurospheres derived from different origins for treatment of rat spinal cord injury.
September 17, 2009 at 7:52 am

Related Articles

Comparison of adult neurospheres derived from different origins for treatment of rat spinal cord injury.

Neurosci Lett. 2009 Jul 24;458(3):116-21

Authors: Zhang HT, Cheng HY, Cai YQ, Ma X, Liu WP, Yan ZJ, Jiang XD, Xu RX

This study is designed to evaluate the therapeutic effects of three types of neurospheres (NSs) derived from brain, bone marrow and adipose tissue in a rat model of spinal contusive injury. As shown by BBB locomotor rating scale and grid test, the optimal therapeutic responses generated by subventricular zone-derived NSs (SVZ-NSs), and followed by adipose-derived (AD-NSs) and bone marrow-derived NSs (BM-NSs) after being grafted into the injured spinal cord. In three cell-treated groups, very few (<1%) grafted cells survived and these survived cells mainly differentiated into oligodendrocytes at week 12 after injury. Additionally, all the cell-treated groups, especially in the SVZ-treated group showed an increase in host oligodendrocytes than control group. Moreover, the level of selective neurotrophins (NTs) in the SVZ-NSs group were significantly higher than those in the BM-NSs and AD-NSs groups, and the level of NTs in the saline group was also significantly higher than sham group. Therefore, not cell replacement or infusion but neuroprotective action associated with endogenous oligodendrocytes and NTs that active by the grafted NSs may contribute to the functional recovery.

PMID: 19394407 [PubMed - indexed for MEDLINE]

 

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