The individual-cell-based cryo-chip for the cryopreservation, manipulation and observation of spatially identifiable cells. I: Methodology

<p>Abstract</p> <p>Background</p> <p>Cryopreservation is the only widely applicable method of storing vital cells for nearly unlimited periods of time. Successful cryopreservation is essential for reproductive medicine, stem cell research, cord blood storage and related biomedical areas. The methods currently used to retrieve a specific cell or a group of individual cells with specific biological properties after cryopreservation are quite complicated and inefficient.</p> <p>Results</p> <p>The present study suggests a new approach in cryopreservation, utilizing the Individual Cell-based Cryo-Chip (i3C). The i3C is made of materials having appropriate durability for cryopreservation conditions. The core of this approach is an array of picowells, each picowell designed to maintain an individual cell during the severe conditions of the freezing - thawing cycle and accompanying treatments. More than 97% of cells were found to retain their position in the picowells throughout the entire freezing - thawing cycle and medium exchange. Thus the comparison between pre-freezing and post-thawing data can be achieved at an individual cell resolution. The intactness of cells undergoing slow freezing and thawing, while residing in the i3C, was found to be similar to that obtained with micro-vials. However, in a fast freezing protocol, the i3C was found to be far superior.</p> <p>Conclusions</p> <p>The results of the present study offer new opportunities for cryopreservation. Using the present methodology, the cryopreservation of individual identifiable cells, and their observation and retrieval, at an individual cell resolution become possible for the first time. This approach facilitates the correlation between cell characteristics before and after the freezing - thawing cycle. Thus, it is expected to significantly enhance current cryopreservation procedures for successful regenerative and reproductive medicine.</p

BIOCOMPATIBILITY OF GLASS-ENCAPSULATED ELECTRONIC CHIPS (TRANSPONDERS) USED FOR THE IDENTIFICATION OF PIGS

The biocompatibility of electronic transponders encapsulated in two different types of glass was studied after they had been implanted subcutaneously into pigs for the purpose of identification. Rods of white crystal glass or green iron-containing glass were screened for superficial impurities by scanning electron microscopy and X-ray analysis, revealing a few crystalline and plaque impurities which were similar for both types of glass, and no differences in elemental composition. In vitro cytotoxicity tests using cell cultures of human dermal fibroblasts and haemolysis and clot formation tests in human blood after contact with the rods, showed that both types of glass were biocompatible. When implanted subcutaneously at the base of the ears of pigs for from three to 150 days, both types of transponder appeared to induce a similar connective tissue capsule, on average less than 0-2 mm in thickness, surrounding the rods. A classic foreign body reaction did not occur. It is concluded that the fibrous capsules were due to scar tissue formed around the glass rods as a result of the tissue being damaged when they were implanted. There were no significant differences between the reactions to the two types of glass. The subcutaneous implantation of glass-encapsulated transponders appears to be a good method for identifying pigs

ANALOGIES IN THE 2-DIMENSIONAL SPATIAL ARRANGEMENT OF ADSORBED PROTEINS AND ADHERING BACTERIA - BOVINE SERUM-ALBUMIN AND STREPTOCOCCUS-SANGUIS-12

Neither proteins nor bacteria adsorb or adhere homogeneously to a substratum surface. The final two-dimensional spatial arrangement depends on a complicated interplay between protein-protein (bacterium-bacterium) and protein (bacterium)-substratum interactions and the prevailing hydrodynamic conditions. In this paper, results are presented of two separate experiments in which bovine serum albumin (BSA) was adsorbed to, or Streptococcus sanguis 12 was deposited on substrata with different wettabilities in a search for analogies in the two-dimensional spatial arrangement of the adsorbed proteins and adhering bacteria. The spatial arrangement of adsorbed BSA, visualized by transmission electron microscopy on replicas of the surface, was island-like on substrata with a low wettability and well distributed on substrata with a high wettability. The spatial arrangement of adhering S. sanguis 12 was observed directly by light microscopy during the experiment and showed a relatively large collection of near-neighbour sites on the low wettability substrata compared with the high wettability substrata. Thus, it seems that the occurrence of island-like structures in protein adsorption is concurrent with a large collection of near-neighbour sites in bacterial adhesion. As a Possible explanation for the above analogy, it is suggested that proteins or bacteria are insufficiently immobilized on low wettability substrata owing to weak interaction forces, and they can move over the substratum surface to yield the two-dimensional spatial arrangements observed