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Cryopreservation of zona-intact porcine embryos has been relatively unsuccessful to date, although some success has been obtained with lipid reduced morulae and early blastocysts. This study adapted some vitrification protocols used successfully with late blastocysts for use with early zona-intact blastocysts, using actin depolymerization, centrifugation,(More)
Cryopreservation nearly universally depends on the equilibration of cells and tissues with high concentrations of permeating chemicals known as cryoprotective agents, or CPAs. Despite their protective properties, CPAs can cause damage as a result of osmotically-driven cell volume changes, as well as chemical toxicity. In this study, we have used previously(More)
Delayed immune reconstitution remains a major cause of morbidity associated with myelosuppression induced by cytotoxic therapy or myeloablative conditioning for stem cell transplant, as well as potentially lethal doses of total- or partial-body irradiation. Restoration of a functional immune cell repertoire requires hematopoietic stem cell reconstitution(More)
The ability to successfully cryopreserve neural cells would represent an important advance with benefits to neural tissue engineering, neural transplantation, and neuroscience research. We have examined key factors responsible for damage to rat embryonic neural cells during cryopreservation using a two-step temperature profile, with an emphasis on the(More)
We have investigated the confounding effects of dynamic range limitations on measurement of the osmotically inactive volume using electrical sensing zone instruments (e.g., Coulter counters), and propose an improved approach to parameter estimation. The conventional approach for analysis of cell size distributions measured by such particle sizing(More)
Coincidence is a phenomenon that occurs when electrical sensing zone instruments fail to temporally resolve two or more particles passing through the sensing zone in close proximity. We have investigated the potential for coincidence errors to confound the estimation of cellular osmotic properties. A mathematical model was developed to predict the magnitude(More)
Granulocytes are currently transfused as soon as possible after collection because they rapidly deteriorate after being removed from the body. This short shelf life complicates the logistics of granulocyte collection, banking, and safety testing. Cryopreservation has the potential to significantly increase shelf life; however, cryopreservation of(More)
We previously measured the membrane water permeability of monolayers and suspensions of MIN6 mouse insulinoma cells at room temperature, and found that water transport was faster in monolayers. Here, we compare water transport kinetics in monolayers and suspensions over a range of temperatures for two different cell types, MIN6 cells and bovine pulmonary(More)
Loading of cryoprotectants into oocytes is an important step of the cryopreservation process, in which the cells are exposed to potentially damaging osmotic stresses and chemical toxicity. Thus, we investigated the use of physics-based mathematical optimization to guide design of cryoprotectant loading methods for mouse and human oocytes. We first examined(More)
The storage of red blood cells (RBCs) in a refrigerated state allows a shelf life of a few weeks, whereas RBCs frozen in 40% glycerol have a shelf life of 10 years. Despite the clear logistical advantages of frozen blood, it is not widely used in transfusion medicine. One of the main reasons is that existing post-thaw washing methods to remove glycerol are(More)