Nejvíce citovaný článek - PubMed ID 30008675
AFM Monitoring the Influence of Selected Cryoprotectants on Regeneration of Cryopreserved Cells Mechanical Properties
Many plants cannot vitrify themselves because they lack glassy state-inducing substances and/or have high water content. Therefore, cryoprotectants are used to induce vitrification. A cryoprotectant must have at least the following primary abilities: high glass-forming property, dehydration strength on a colligative basis to dehydrate plant cells to induce the vitrification state, and must not be toxic for plants. This review introduces the compounds used for vitrification solutions (VSs), their properties indicating a modification of different plant vitrification solutions, their modifications in the compounds, and/or their concentration. An experimental comparison is listed based on the survival or regeneration rate of one particular species after using more than three different VSs or their modifications. A brief overview of various cryopreservation methods using the Plant Vitrification Solution (PVS) is also included. This review can help in alert researchers to newly introduced PVSs for plant vitrification cryoprotocols, their properties, and the choice of their modifications in the compounds and/or their concentration.
- Klíčová slova
- cryoprotectant, glassy state, toxicity, ultra-low temperature,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Spinocerebellar ataxia type-1 (SCA1) is caused by an abnormally expanded polyglutamine (polyQ) tract in ataxin-1. These expansions are responsible for protein misfolding and self-assembly into intranuclear inclusion bodies (IIBs) that are somehow linked to neuronal death. However, owing to lack of a suitable cellular model, the downstream consequences of IIB formation are yet to be resolved. Here, we describe a nuclear protein aggregation model of pathogenic human ataxin-1 and characterize IIB effects. Using an inducible Sleeping Beauty transposon system, we overexpressed the ATXN1(Q82) gene in human mesenchymal stem cells that are resistant to the early cytotoxic effects caused by the expression of the mutant protein. We characterized the structure and the protein composition of insoluble polyQ IIBs which gradually occupy the nuclei and are responsible for the generation of reactive oxygen species. In response to their formation, our transcriptome analysis reveals a cerebellum-specific perturbed protein interaction network, primarily affecting protein synthesis. We propose that insoluble polyQ IIBs cause oxidative and nucleolar stress and affect the assembly of the ribosome by capturing or down-regulating essential components. The inducible cell system can be utilized to decipher the cellular consequences of polyQ protein aggregation. Our strategy provides a broadly applicable methodology for studying polyQ diseases.
- Klíčová slova
- Ataxin-1, Oxidative stress, Polyglutamine, Protein network, Ribosome, Sleeping beauty transposon,
- MeSH
- ataxin-1 genetika metabolismus MeSH
- intranukleární inkluzní tělíska * metabolismus MeSH
- jaderné proteiny genetika metabolismus MeSH
- lidé MeSH
- oxidační stres MeSH
- proteiny nervové tkáně * genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- ataxin-1 MeSH
- jaderné proteiny MeSH
- proteiny nervové tkáně * MeSH
In our work, we developed the synthesis of new polyfunctional pegylated trehalose derivatives and evaluated their cryoprotective effect using flow cytometry. We showed that new compounds (modified trehaloses) bound to appropriate extracellular polymeric cryoprotectants could be helpful as a chemical tool for the evaluation of their potential toxic cell membrane influences. Our aim was to form a chemical tool for the evaluation of cryoprotectant cell membrane influences, which are still not easily predicted during the freezing/thawing process. We combined two basic cryoprotectants: polyethyleneglycols (PEGs) and trehalose in the new chemical compounds-pegylated trehalose hybrids. If PEG and trehalose are chemically bound and trehalose is adsorbed on the cell surface PEGs molecules which are, due to the chemical bonding with trehalose, close to the cell surface, can remove the cell surface hydration layer which destabilizes the cell membrane. This was confirmed by the comparison of new material, PEG, trehalose, and their mixture cryoprotective capabilities.
- Klíčová slova
- click-chemistry, cryoprotection, pegylation,
- MeSH
- buněčná membrána účinky léků metabolismus MeSH
- dimethylsulfoxid farmakologie MeSH
- kryoprezervace MeSH
- kryoprotektivní látky farmakologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- polyethylenglykoly chemie farmakologie MeSH
- průtoková cytometrie MeSH
- syntetická chemie okamžité shody MeSH
- trehalosa analogy a deriváty chemická syntéza chemie farmakologie MeSH
- viabilita buněk účinky léků MeSH
- zmrazování MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dimethylsulfoxid MeSH
- kryoprotektivní látky MeSH
- polyethylenglykoly MeSH
- trehalosa MeSH