Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to ...

Preservation methods on the physiological and brewing technical characters in bottom and top brewing yeast strains were investigated. The preserved yeasts were reactivated after 24 months storage and grown up to stationary phase. The samples of filter paper storage indicated a higher cell growth and viability during propagation than those ...

The natural nutrient component Curcumin with anti-inflammatory and antitumor activity has previously been shown to stimulate apoptosis of several nucleated cell types. The present study has been performed to explore whether Curcumin could similarly induce suicidal death of erythrocytes or eryptosis, which is characterized by cell shrinkage and cell membrane ...

Seasonal changes in the accumulation of soluble sugars in extracellular freezing cortical parenchyma cells and deep supercooling xylem parenchyma cells in Japanese white birch (Betula platyphylla var. japonica) were compared to identify the effects of soluble sugars on the mechanism of deep supercooling, which keeps the liquid state of water ...

We investigated whether the addition of Gingko Biloba extract (EGb 761) to rabbit corneal epithelial medium before cell freezing improved cell viability after freezing then thawing. After removal of corneas, they were treated with enzymes and the corneal epithelium was prepared as a single cell suspension in freezing media with ...

The long-term storage of human and animal cell lines is a relatively new branch of the science of cryopreservation of living organisms. The need to maintain the increasing numbers of cell lines, as they started to emerge from research laboratories from the mid-1900s, necessitated a radical approach to the problem. ...

Successful cryopreservation of plant shoot tips is dependent upon effective desiccation through osmotic or physical processes. Microscopy techniques were used to determine the extent of cellular damage and plasmolysis that occurs in peppermint (Mentha x piperita) shoot tips during the process of cryopreservation, using the cryoprotectant plant vitrification solution 2 ...

Here we describe a protocol for the production of frozen competent yeast cells that can be transformed with high efficiency using the lithium acetate/single-stranded carrier DNA/PEG method. This protocol allows the production of highly competent yeast cells that can be frozen and used at a later date and is especially ...

Hemoglobins are ancient O(2)-binding proteins, ubiquitously found in eukaryotes. They have been categorized as symbiotic, nonsymbiotic and truncated hemoglobins. We have investigated the cellular localization of nonsymbiotic hemoglobin proteins during somatic embryogenesis in Cichorium hybrid leaves (Cichorium intybus L. var. sativum x C. endivia var. latifolia) using immunolocalization technique. These ...

Freeze storage of human embryonic stem (hES) cells has not proven effective using the methods employed for mouse ES (mES) cells, while rhesus ES (rhES) cells are only modestly effectively frozen using common mES cell methods. Because human and rhES cells are passaged and frozen in clusters that approximate the ...

The aim of this study was to investigate the effect of whey protein isolate (WPI) gel microentrapment on the viability of Lactobacillus rhamnosus R011 during the production and storage of biscuits, frozen cranberry juice, and vegetable juice. Viability of microentrapped (ME) cells was compared to free cells freeze-dried in a ...

The deformability of erythrocytes is of great importance for oxygen delivery in the microcirculation [Lipowsky, H.H., 2005. Microvascular rheology and hemodynamics. Microcirculation 12, 5-15]. Aging of erythrocytes is associated with a reduction in deformability and also in size. The present work describes an automated cell analyzer which utilizes a glass ...

A major challenge in the widespread application of hES (human embryonic stem) cells in clinical therapy and basic scientific research is the development of efficient cryopreservation protocols. Conventional slow-cooling protocols utilizing standard cryoprotectant concentrations i.e. 10% (v/v) DMSO, yield extremely low survival rates of less than 5% as reported by ...

Lead and cadmium content of different brands and sizes (N=38) of dry cell batteries (zinc-carbon Leclanche systems) available in Nigeria were determined by atomic absorption spectrometry after acid digestion. The average Pb and Cd content of the dry cells are 1077.9±751.7mg/kg (Range 42-3170mg/kg) and 108.3±81.4mg/kg (Range 4.6-410mg/kg) respectively. The mean ...

Here we demonstrate how our lab freezes HuES human embryonic stem cell lines. A healthy, exponentially expanding culture is washed with PBS to remove residual media that could otherwise quench the Trypsin reaction. Warmed 0.05% Trypsin-EDTA is then added to cover the cells, and the plate allowed to incubate for ...

The cryoprotective effect of intracellular free high-mannose oligosaccharides (HMOS) on mammalian cells and proteins was examined by monitoring PC-12 cell viability and assaying protein kinase C (PKC)-epsilon activity. 1-Deoxymannojirimycin, an inhibitor of alpha-mannosidase, to cause an increase in intracellular free HMOS, significantly rescued PC-12 cells with 2-h freezing insult at ...

Transgenic suspension cells of Oryza sativa L. cv. Dongjin utilized as a host for producing recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) were preserved in liquid nitrogen (-196 degrees C) after slow prefreezing in a deep freezer (-70 degrees C). The development of an optimal procedure for long-term storage was ...

This study established a method for preserving chicken primordial germ cells (PGC) that enables long-term storage in liquid N. Gonads were harvested from stage 27 chick embryos and pooled in groups of 5, 10 (10E), or 20 embryos, contributing gonads to the cell suspension. The gonadal cells, including PGC, were ...

The determinants of sickle red blood cell (RBC) life span have not been well-defined but may include both intrinsic factors (eg, the tendency to sickle) and extrinsic factors (eg, the capacity of the reticuloendothelial system to remove defective RBCs). Fetal hemoglobin (HbF) is heterogeneously distributed among sickle RBCs; F cells ...

Current knowledge on the pathophysiology of sickle cell anemia is reviewed and potential therapeutic options are reviewed and discussed. It is hoped that better understanding of the pathophysiology will improve the care given to these patients as well as their quality of life. <BR><BR> Keywords : sickle cell anemia, pathophysiology, ...

We tested a "standard" cryopreservation protocol (slow cooling with 10% DMSO) on the human embryonic stem cell (hESC) line H9 containing an Oct-4 (POU5F1) promoter-driven, enhanced green fluorescent protein (EGFP) reporter to monitor maintenance of pluripotency. Cells were cooled to -80 degrees C in cryovials and then transferred to a ...

In the forensic science laboratory, the recovery of spermatozoa from vaginal swabs, or vaginal cells from penile swabs, can help determine if sexual intercourse may have taken place. There are several methods used to recover spermatozoa and cells from the swabs before visualisation on a microscope slide and most of ...

Two pennate microphytobenthic diatoms, Amphora coffeaeformis (Agardh) Kutzing and Navicula transitans var. derasa f. delicatula Heimdal, were cryopreserved and monitored on thawing to track the mechanical injuries and their post-preservation recovery. Cells were subjected to (1) direct freezing in liquid nitrogen and (2) two-step cooling with and without the cryoprotectant, ...

We have further characterized villous trophoblasts isolated by trypsinization and purified by elimination of CD9/MHC class I/MHC class II expressing cells. The cells isolated were >99.99% cytotrophoblasts by criteria of cytokeratin (positive) and vimentin (negative) expression. Purified cells directly after isolation (fresh) were compared with cryopreserved and thawed cells (frozen) ...

BACKGROUND: Long-term storage of platelets (PLTs) in the dry state would greatly improve options for PLT storage. Whether trehalose-loaded freeze-dried and rehydrated PLTs could regulate intracellular pH (pHi) was evaluated. STUDY DESIGN AND METHODS: Previously it was shown that human PLTs can be successfully preserved by freeze-drying with trehalose. Trehalose-loaded ...

Cryopreservation is widely used for long-term conservation of various tissues, embryos or gametes. However, few studies have described cryopreservation of invertebrate primary cell cultures and more particularly of marine invertebrate somatic cells. This technique would however be of great interest to facilitate the study of various metabolic processes which vary ...

The preservation of micro-organisms by different drying methodologies has been used for decades. Freeze drying in particular is the preferred method for transporting and storing vast culture collections of micro-organism strain types. The literature on drying and preserving micro-organisms is extensive, but is often specific to one particular strain. This ...

The purpose of this study was to test the authors' hypothesis that previously cryopreserved adipose aspirates collected from conventional liposuction could still be a reliable source of human processed lipoaspirate cells. Adipose aspirates were collected from 12 adult female patients after conventional liposuction of the abdomen and were then preserved ...

The mechanism of cell injury during slow freezing was examined using PC-3 human prostate adenocarcinoma cells suspended in NaCl solutions. The objective was to evaluate contribution of extracellular ice and the 'solution effects' to freezing injury separately. The solution effects that designate the influence of elevated concentration were evaluated from ...

One of the stumbling blocks in the formation of a thermoresponsive cell-hydrogel hybrid (TCH) is the efficient incorporation of cells in thermoresponsive hydrogels (TH) using traditional top-down (i.e., cells penetrate in the pre-set gels from top surface) approach. This approach is slow and tedious because the hydrogel needs to soak ...

Mosses are known to have the ability to develop high degrees of resistance to desiccation and freezing stress at cellular levels. However, underlying cellular mechanisms leading to the development of stress resistance in mosses are not understood. We previously showed that freezing tolerance in protonema cells of the moss Physcomitrella ...

High-pressure freezing avoids the artefacts induced by conventional chemical fixation, and, in combination with freeze-substitution and plastic embedding, is a reliable method for the ultrastructural analysis of mammalian cell monolayers. In order to high-pressure freeze mammalian cell monolayers, cells have to be seeded on a suitable substrate. Unfortunately, electron microscopy ...

A combination of gene and cell-based therapies may provide significant advantages over existing treatments in terms of their effectiveness. However, long-term efficient gene delivery has been difficult to achieve in many cell types, including endothelial cells. We developed a freeze-thaw technique which significantly increases the transduction efficiency of recombinant adeno-associated ...

AIMS: To assess four carbohydrates for the protective effect against Lactobacillus helveticus cells inactivation during vacuum drying, and to study the effect of selected carbohydrate on changes of inactivation kinetics. METHODS AND RESULTS: Early stationary phase L. helveticus cells grown in MRS media were recovered from fermentation broth, washed with ...

The aim of the present study was to test the use of cryopreserved cells for human ether-ago- go (hERG) channel screening. A stable Chinese hamster ovary-K1 cell line expressing hERG channels was used in a non-radioactive Rb+ efflux assay for screening hERG channel activity. Cells prepared from normal cultures and ...

BACKGROUND: Cold ischemia and reperfusion during renal transplantation result in release of reactive oxygen species. The aim of this study is to examine whether cold storage induced cell injury can be ameliorated by adding flavonoids directly to preservation solutions. METHODS: Cultured renal tubular epithelial cells (LLC-PK1) were stored in University ...

Chorispora bungeana Fisch. & C.A. Mey (Crucifer) is a rare alpine subnival plant surviving sudden snowstorms. In this paper, we have attempted to explore possible roles of autoxidation rate (AR) and the antioxidant enzymes associated with cryoprotective mechanisms in the plant cells. The results showed that when the suspension cultures ...

International Dairy Journal Vol.16 Nr.12, 1477 - 1482

The clinical implementation of gene therapy requires large-scale production of viral vector stocks (VS) derived from packaging cell lines. Upon scaling-up, maintenance of high viral titers and filtration of the VS become significantly challenging. Thus, production schemes amenable to straightforward validation must be developed. To this end, we have established ...

Sucrose has been reported to play multiple roles in the winter biology of temperate woody species. However, no report on the molecular basis of sucrose transport in xylem tissue has yet been made. In the walnut tree, it is demonstrated that during the autumn-winter period, active absorption of sucrose from ...

Encapsulation-vitrification, which is a combination of encapsulation-dehydration and vitrification procedures, is a newly developed technique for cryopreservation of plant germoplasm. Here, we describe the protocol of this methodology, using grapevine (Vitis) as a model plant. Cell suspensions at the exponential growth stage were encapsulated with 2.5% sodium alginate solution in ...

Factors affecting cell viability, plating efficiency, and survival of hepatocytes after cryopreservation have been investigated. We focused especially on the effect of including trehalose and related oligosaccharides in the cryopreservation fluid. This was supplemented with either glucose, trehalose, maltotriose, or other sugars, in addition to dimethyl sulfoxide (10%) and first ...

A major challenge in the widespread application of human embryonic stem (hES) cells in clinical therapy and basic scientific research is the development of efficient cryopreservation protocols. Conventional slow-cooling protocols utilizing standard cryoprotectant concentrations i.e. 10% (v/v) DMSO, yield extremely low survival rates of <5% as reported by previous studies. ...

Evidence has accumulated that periosteal cells have a great potential to regenerate bone. We have demonstrated that cultured periosteum (CP) in membrane form is an effective device to regenerate alveolar bone. To increase the availability of CP in a clinical environment, an effective cryopreservation protocol for CP has been developed. ...

Understanding the biophysical processes that govern freezing injury of a tissue equivalent (TE) is an important step in characterizing and improving the cryopreservation of these systems. TEs were formed by entrapping human dermal fibroblasts (HDFs) in collagen or in fibrin gels. Freezing studies were conducted using a Linkam cryostage fitted ...

Understanding the ultrastructural response of cells to the freezing process is important for designing cryopreservation strategies for cells and tissues. The cellular structures of attached cells are targets of cryopreservation-induced damage. Specific fluorescence staining was used to assess the status of the actin filaments (F-actin) of murine osteoblasts attached to ...

Eggshell membranes (ESM) have been shown to exhibit antibacterial activity. The purpose of this study was to evaluate the enzymatic and biological [decimal reduction times (D-values)] activities of ESM as a function of bird breed, age, and ESM stabilization treatments. Younger White Leghorn (WL) hens produced ESM with 28% higher ...

A study was done to determine the performance of differential, selective media for supporting resuscitation and colony development by stressed cells of Enterobacter sakazakii. Cells of four strains of E. sakazakii isolated from powdered infant formula were exposed to five stress conditions: heat (55 degrees C for 5 min), freezing ...

Mammalian cells appear to be naturally tolerant to cold temperatures, but the formation of ice when cells are cooled leads to a variety of damaging effects. The study of cryo-injury, therefore, becomes the study of when and how ice is formed both inside and outside the cell during cooling. Protectant ...

To produce commercial biocontrol agents (BCAs) successfully, it is important that cheap and economic substrates are used which support high numbers of good quality inoculum. Production of formulations conserving ecological competence and shelf-life should also be ensured. With this in mind, studies focusing on yeast ecophysiology were conducted to produce ...