Liquidus Tracking: Controlled Rate Vitrification for the Cryopreservation of Larger Volumes and Tissues

BACKGROUND: Vitrification of cells or tissue at controlled cooling rates suitable for larger volumes, and with reduced cryoprotectant toxicity. OBJECTIVE: To set out the current understanding of the LiquidusTracking (LT) vitrification technique, and to discuss the challenges and benefits of translating the method into laboratory protocols more generally applicable to meet requirements of large volume and 3-D cryo-banking in the era of regenerative medicine. METHODS: By adding small amounts of cryoprotectants at each step and subsequently cooling the sample just above its freezing point before further increasing CPA concentration, cryoprotectant toxicity is minimized. RESULT: CPA toxicity can be reduced by lowering the temperature. Different manual approaches to LT were evaluated and further improved. CONCLUSIONS: Manual liquidus tracking is complicated and exhibits potential high variability. Nevertheless, this approach offers the possibility of testing several conditions simultaneously and could be used to pre-test conditions prior to automatic LT development

On Two Models of the Light Pulse Delay in a Saturable Absorber

A comparative analysis of two approaches to description of the light modulation pulse delay in a saturable absorber is presented. According to the simplest model, the delay of the optical pulse is a result of distortion of its shape due to absorption self-modulation in the nonlinear medium. The second model of the effect, proposed at the beginning of our century, connects the pulse delay with the so-called "slow light" resulting from the group velocity reduction under conditions of the coherent population oscillations. It is shown that all the known experimental data on the light pulse delay in saturable absorbers can be comprehensively described in the framework of the simplest model of saturable absorber and do not require invoking the effect of coherent population oscillations with spectral hole-burning and anomalous modifications of the light group velocity. It is concluded that the effect of group velocity reduction under conditions of coherent population oscillations has not received so far any experimental confirmation, and the assertions about real observation of the "slow light" based on this mechanism are groundless.Comment: Regretfully, the journal version of the paper (in Optics and Spectroscopy) appeared to be strongly corrupted due to ignorant editing. In particular, "coherent population oscillations" (CPO) was replaced by "population coherent oscillations" (PCO), "bleaching" - by "clearing", and "bleachable absorber " - by "clearable absorber". Here we present original version of the pape

Report on a collecting trip of the British Myriapod Group to Hungary in 1994

During a collecting trip participated jointly by the members of the British Myriapod Group and by Hungarian experts in 1994, 34 species of millipedes, 14 of centipedes, 8 of woodlice and 73 of spiders were recorded from Hungary. Two records of the millipede species Boreoiulus tenuis (Bigler, 1913) and Styrioiulus styricus (Verhoeff, 1896) were new to the fauna of Hungary

Lithic Morphological Organization: Gahagan Bifaces from Texas and Louisiana

This study is focused upon an analysis of Gahagan biface morphology and enlists the three largest samples of these bifaces, to include that of the type site (Gahagan Mound) as well as the Mounds Plantation and George C. Davis sites. Results indicate a significant difference in Gahagan biface morphology at the Mounds Plantation site when compared with Gahagan bifaces from the Gahagan Mound and George C. Davis sites. A test of morphological integration indicates that the bifaces are significantly integrated, meaning that those traits used to characterize their shape (blade and base) vary in a coordinated manner. Tests for allometry and asymmetry were not significant. Results confirm that Gahagan biface production at Mounds Plantation differs significantly when compared with industries at Gahagan Mound and George C. Davis. Results augment previous inquiries, and provide additional evidence for a north-south divide based upon the morphology associated with communities of practice for Gahagan bifaces. When viewed in concert with similar shifts in Hickory Fine Engraved and Smithport Plain bottle morphology, multiple lines of evidence lend support to an increasingly robust argument for two previously unrecognized and morphologically-unique Caddo communities of practice

Small-Scale Fluidized Bed Bioreactor for Long-Term Dynamic Culture of 3D Cell Constructs and in vitro Testing

With the increasing interest in three-dimensional (3D) cell constructs that better represent native tissues, comes the need to also invest in devices, i.e., bioreactors, that provide a controlled dynamic environment similar to the perfusion mechanism observed in vivo. Here a laboratory-scale fluidized bed bioreactor (sFBB) was designed for hydrogel (i.e., alginate) encapsulated cells to generate a dynamic culture system that produced a homogenous milieu and host substantial biomass for long-term evolution of tissue-like structures and “per cell” performance analysis. The bioreactor design, conceptualized through scale-down empirical similarity rules, was initially validated through computational fluid dynamics analysis for the distributor capacity of homogenously dispersing the flow with an average fluid velocity of 4.596 × 10–4 m/s. Experimental tests then demonstrated a consistent fluidization of hydrogel spheres, while maintaining shape and integrity (606.9 ± 99.3 μm diameter and 0.96 shape factor). It also induced mass transfer in and out of the hydrogel at a faster rate than static conditions. Finally, the sFBB sustained culture of alginate encapsulated hepatoblastoma cells for 12 days promoting proliferation into highly viable (>97%) cell spheroids at a high final density of 27.3 ± 0.78 million cells/mL beads. This was reproducible across multiple units set up in parallel and operating simultaneously. The sFBB prototype constitutes a simple and robust tool to generate 3D cell constructs, expandable into a multi-unit setup for simultaneous observations and for future development and biological evaluation of in vitro tissue models and their responses to different agents, increasing the complexity and speed of R&D processes

Experimental and Computational Study of Area and Perimeter Contributions to Radiometer Forces

The relative contribution to the radiometric force of the area and perimeter of the vane is studied experimentally and numerically. Experimentally, a circular vane, a low-aspect rectangular vane, and a high-aspect rectangular vane were all tested on a force balance, with nano-Newton resolution, placed in a stagnant gas. The computational results were obtained through 2-D simulations using the direct simulation Monte Carlo method, as well as a discrete ordinate solution of the ES model kinetic equation. Gas pressure was varied from 0.006 to 6 Pa, which was a broad enough range to observe the characteristic peak force production of a radiometer in the transition regime, where the peak occurs at Kn ~ 0.1. It was found that the area of a radiometer vane is responsible for a significant amount ofthe total force production through a wide range of operating pressures. It is only at the highest background pressures, well after force production has peaked, that the vane perimeter appears to dominate the operation of the radiometer

Decisional tool for cost of goods analysis of bioartificial liver devices for routine clinical use

BACKGROUND AIMS: Bioartificial liver devices (BALs) are categorized as advanced therapy medicinal products (ATMPs) with the potential to provide temporary liver support for liver failure patients. However, to meet commercial demands, next-generation BAL manufacturing processes need to be designed that are scalable and financially feasible. The authors describe the development and application of a process economics decisional tool to determine the cost of goods (COG) of alternative BAL process flowsheets across a range of industrial scales. METHODS: The decisional tool comprised an information database linked to a process economics engine, with equipment sizing, resource consumption, capital investment and COG calculations for the whole bioprocess, from cell expansion and encapsulation to fluidized bed bioreactor (FBB) culture to cryopreservation and cryorecovery. Four different flowsheet configurations were evaluated across demands, with cell factories or microcarriers in suspension culture for the cell expansion step and single-use or stainless steel technology for the FBB culture step. RESULTS: The tool outputs demonstrated that the lowest COG was achieved with microcarriers and stainless steel technology independent of the annual demand (1500-30 000 BALs/year). The analysis identified the key cost drivers were parameters impacting the medium volume and cost. CONCLUSIONS: The tool outputs can be used to identify cost-effective and scalable bioprocesses early in the development process and minimize the risk of failing to meet commercial demands due to technology choices. The tool predictions serve as a useful benchmark for manufacturing ATMPs

Applications and optimization of cryopreservation technologies to cellular therapeutics

Delivery of cell therapies often requires the ability to hold products in readiness whilst logistical, regulatory and potency considerations are dealt with and recorded. This requires reversibly stopping biological time, a process which is often achieved by cryopreservation. However, cryopreservation itself poses many biological and biophysical challenges to living cells that need to be understood in order to apply the low temperature technologies to their best advantage. This review sets out the history of applied cryopreservation, our current understanding of the various processes involved in storage at cryogenic temperatures, and challenges for robust and reliable uses of cryopreservation within the cell therapy arena