Never Say Die

This chapter evaluates the longevity industry and the claims being made about the viability of radical life extension. It distinguishes between the established science of human aging and the emerging promissory economy being built upon visions of extending life well beyond the theoretical optimal human lifespan of approximately 115 years. With reference to examples such as cryo-preservation and cellular interventions, this wide-ranging exploration of the contemporary Western obsession with prolonging vibrant life focuses on the techno-political dimensions of “disrupting death” and the complex relationship between hope and hype that sustains the immortality imaginary.

Tasa de preñez post vitrificación en los embriones de équidos producidos in vivo. Revisión

Vitrification is a cryo-preservation method often used in embryos obtained from mares or jennies. It consists in the dramatic reduction of temperature to levels close to -196 °C, that allows the cryopreserving solution containing the embryo to pass from liquid to vitreous state. Several improvements to vitrification protocols have made possible to cryo-preserve embryos with different sizes; since during the first decade after the year 2000, only small embryos were successfully vitrified. Embryos collected at the sixth day post ovulation (PO) are usually smaller or equal to 300 micrometers in diameter (≤ 300 µmØ) and can be routinely vitrified following simple protocols; they have a higher post vitrification pregnancy rate (PVPR) when compared to large embryos which have more than 300 micrometers in diameter (˃ 300 µmØ). The high PVPR of embryos ≤ 300 µmØ is due to an embryo capsule (EC) that is not fully developed yet and has a high permeability to cryo-preserving solutions. At present time, embryos collected either the seventh or eighth day PO are ˃ 300 µmØ and are characterized to have a low post vitrification survival; in order to increase their PVPR their EC might be punctured to make it permeable to cryopreserving solutions. Additionally, there are at least two factors that can be manipulated to increase the PVPR of embryos ˃ 300 µmØ; one is to reduce their size by aspiring their blastocoelic liquid (BL), and the other is to induce a high temperature transfer index (TTI) to rapidly reach -196 °C.

Influence cryo preservation on viability of Chamomilla reticuta’ seeds varia «Podmoskovnaya» and chemical composition of essential oil

The article examines the effect above critical low temperatures on the quantitative and qualitative composition of essential oil of Chamomilla reticuta varia «Podmoskovnaya». Seeds of the studied plant species were treated with ultra-low temperature (–196 ºC) before planting in open ground and the viability of the seeds was determined under laboratory conditions. Cryo processing was carried out by direct immersion of seeds in plastic tubes in Dewar vessels with liquid nitrogen. Cryoprotectants were not used. Cryo processing duration was 1 hour, 3 hour and step freezing. Step freezing of seeds was performed in the following sequence: 1 hour in the refrigerating compartment (+ 4 ºC), 1 hour in the freezing compartment (–8 ºC) and 1 hour in liquid nitrogen. In all cases of the experiment, a slow thawing regime was applied. After cryogenic exposure, the material was planted into open ground. Recovery of essential oil was carried out by hydro distillation on a Clevenger apparatus. The component composition of the essential oils was determined on a Clarus-SQ 8 gas chromatograph with a mass spectrometric detector. As a result of the study, it was found that the effect of ultra-low temperatures on the seed material of the Moscow Region chamomile did not negatively affect the quantitative and qualitative composition of the essential oil of the pharmacy chamomile. On the contrary, this method of treating seeds has had a positive effect on the quantitative composition of the essential oil during the growth of the plant and in some cases has even increased the percentage of certain components, such as αfarnesene, β-farnesene, spatulenol, cis-en-in-dicycloether.

Seed germination of Allochrusa gypsophiloides(Caryophyllaceae), an endemic species from Central Asia and Kazakhstan

The results of evaluating the laboratory seed germination of endemic Allochrusa gypsophiloides (Turkestan soap root), depending on storage conditions in combination with gibberellic acid treatment (GA3), are presented. In dry storage, control seeds were characterised by a long after-ripening period and a fluctuating germination behaviour upon removal from storage, with a maximum value of 23%. The sensitivity of seeds to GA3 during dry storage varied significantly, with two germination peaks at 5-7 months, and 12 months (37.5 and 50% germination, respectively). Cold stratification and cryo-preservation accelerated seed after-ripening, promoted germination synchronisation and increased seed sensitivity to GA3. The cold stratification of seeds increased germination four months earlier than during dry storage. GA3 increased germination from 16.7 and 18.3% for the control to 37.5 and 45% for seeds cryopreserved for 5 and 12 months, respectively. We recommend cryopreserving Turkestan soap root seeds to avoid viability loss and to then germinate the seeds after pretreatment with GA3.

CryoSIM: super resolution 3D structured illumination cryogenic fluorescence microscopy for correlated ultra-structural imaging

Rapid cryo-preservation of biological specimens is the gold standard for visualising cellular structures in their true structural context. However, current commercial cryo-fluorescence microscopes are limited to low resolutions. To fill this gap, we have developed cryoSIM, a microscope for 3D super-resolution fluorescence cryo-imaging for correlation with cryo electron microscopy or cryo soft X-ray tomography. We provide the full instructions for replicating the instrument mostly from off-the-shelf components and accessible, user-friendly open source Python control software. Therefore, cryoSIM democratises the ability to detect molecules using super-resolution fluorescence imaging of cryo-preserved specimens for correlation with their cellular ultrastructure.

Correlative cryo-structured illumination fluorescence microscopy and soft X-ray tomography elucidates reovirus intracellular release pathway

Imaging of biological matter across resolution scales presents the challenge of preserving the direct and unambiguous correlation of subject features from the macroscopic to the microscopic level. We present here a correlative imaging platform developed specifically for imaging cells in 3D, under cryogenic conditions. Rapid cryo-preservation of biological specimens is the current gold standard in sample preparation for ultrastructural analysis in X-ray imaging. However, cryogenic fluorescence localisation methods are by and large diffraction-limited and fail to deliver matching resolution. We addressed this technological gap by developing an integrated, user-friendly, platform for 3D correlative imaging of cells in cryo-preserved states using super-resolution structured illumination microscopy (SIM) in conjunction with soft X-ray tomography (SXT). The power of this new approach is demonstrated by studying the process of reovirus release from intracellular vesicles during the early stages of infection and identifying novel virus-induced structures.

Emerging Technologies in Scar Management: The Role of Allogeneic Cells

Scars caused by burns, chronic ulcers from diabetes, infections, skin cancer surgery, and other genetic or somatic disease could require effective treatment to avoid functional and psychological troubles and even mortality. Most of the current treatments aim to reduce local inflammation but not to prevent scarring. Herein, we discussed about emerging technologies in scar management using allogeneic cell therapy. The industrialised allogeneic cell therapy products and the clinical trials using keratinocytes, fibroblasts or MSCs demonstrated acceleration of skin cell migration and proliferation, control wound scarring, immunomodulatory properties and improved angiogenesis. In addition, allogeneic cell transplants offered the possibility of large pre-fabrication, cryo-preservation, for instantaneous use and repeated applications. Current research exploring allogeneic cell therapies for scar treatment are focusing on grafting of epidermal sheets, cellular dermal substitutes and reconstructed skin equivalent and cell intradermal injections. Advances in knowledge in therapeutic potentials of allogeneic injected cells give rise to new therapeutic approaches such as administration of allogeneic cell-derived extracellular vesicles.

CRYO PRESERVATION OF HUMAN DENDRITIC CELLS FOR CLINICAL USE

Introduction. The increasing clinical use of cellular technologies suggests the possibility of long-term storage of the cellular product while maintaining its viability and basic properties. The procedures of cell’s cryopreservation used in laboratory as well in clinical practice differ a lot. Each method includes two tasks to solve: what is the optimal freezing medium to use and what cryopreservation procedure to prefer. In this paper, we present the method utilized in our center for bone marrow cell cryopreservation. The freezing was carried out in nitrogen vapor after adding the medium containing 95 % dextran [average mw 50 000–70 000] and 5 % dimethylsulfoxid.Purpose. To show that the proposed method of cryopreservation of dendritic cells is highly effective, simple, reproducible and most convenient for clinical use.Materials and methods. Viability, expression of surface antigens and stimulating activity towards allogeneic T lymphocytes of cryopreserved mature dendritic cells cultured from peripheral blood monocytes were evaluated.Results. The first cryopreservation resulted in the death of a small amount of cells. The second freezing procedure increased the proportion of dead cells. Meanwhile, the difference in the expression of the surface antigens in fresh, cryopreserved and re-cryopreserved dendritic cells was not statistically significant. The level of stimulating activity of fresh and cryopreserved dendritic cells did not significantly differ. Conversely the proliferation of allogeneic T lymphocytes was decreased after stimulation with re-cryopreserved dendritic cells.Conclusion. The presented method of cryopreservation allows to preserve the viability and basic functions of dendritic cells. After thawing dendritic vaccine could be administered to patients after being diluted in an isotonic saline without washing, which makes this method the most convenient for clinical use.