Long-Term Cryopreservation Does Not Affect Quality of Peripheral Blood Stem Cell Grafts: A Comparative Study of Native, Short-Term and Long-Term Cryopreserved Haematopoietic Stem Cells.

Cryopreserved haematopoietic progenitor cells are used to restore autologous haematopoiesis after high dose chemotherapy. Although the cells are routinely stored for a long period, concerns remain about the maximum storage time and the possible negative effect of storage on their potency. We evaluated the effect of cryopreservation on the quality of peripheral stem cell grafts stored for a short (3 months) and a long (10 years) period and we compared it to native products.The viability of CD34+ cells remained unaffected during storage, the apoptotic cells were represented up to 10% and did not differ between groups. The clonogenic activity measured by ATP production has decreased with the length of storage (ATP/cell 1.28 nM in native vs. 0.63 in long term stored products, P < 0.05). Only borderline changes without statistical significance were detected when examining mitochondrial and aldehyde dehydrogenase metabolic activity and intracellular pH, showing their good preservation during cell storage. Our experience demonstrates that cryostorage has no major negative effect on stem cell quality and potency, and therefore autologous stem cells can be stored safely for an extended period of at least 10 years. On the other hand, long term storage for 10 years and longer may lead to mild reduction of clonogenic capacity. When a sufficient dose of stem cells is infused, these changes will not have a clinical impact. However, in products stored beyond 10 years, especially when a low number of CD34+ cells is available, the quality of stem cell graft should be verified before infusion using the appropriate potency assays.

Immunological aspects of COVID-19: What do we know?

The newly emerged coronavirus (severe acute respiratory syndrome coronavirus 2 SARS-CoV-2) and the disease that it causes coronavirus disease 2019 (COVID-19) have changed the world we know. Yet, the origin and evolution of SARS-CoV-2 remain mostly vague. Many virulence factors and immune mechanisms contribute to the deteriorating effects on the organism during SARS-CoV-2 infection. Both humoral and cellular immune responses are involved in the pathophysiology of the disease, where the principal and effective immune response towards viral infection is the cell-mediated immunity. The clinical picture of COVID-19, which includes immune memory and reinfection, remains unclear and unpredictable. However, many hopes are put in developing an effective vaccine against the virus, and different therapeutic options have been implemented to find effective, even though not specific, treatment to the disease. We can assume that the interaction between the SARS-CoV-2 virus and the individual's immune system determines the onset and development of the disease significantly.

Cryopreserved NK cells in the treatment of haematological malignancies: preclinical study.

BACKGROUND: Leukaemia is an aggressive cancer of haematopoiesis. Despite increasing treatment success, the relapse rate is still high. Natural killer (NK) cells play a key role in the immune response to malignancies; thus, it is conceivable that NK cell-based immunotherapy may control relapses, while extending the disease-free survival. In our study, we investigated whether cryopreserved NK cells are able to kill the leukaemic K562 cell line, the necessity of IL-2 co-application and the association of activation marker expression (NKp44, NKG2D and CD25) with cytotoxic potential. MATERIALS AND METHODS: K562 cells were added to NK cell cultures in different ratios, i.e. 1:5, 1:10 and 1:20 (K562/NK), immediately after thawing NK cells or after 3-6-12-24 h of re-cultivation with or without IL-2. RESULTS: Our results demonstrated the ability of cryopreserved NK cells to kill K562 in all ratios, times and culture conditions. The number of dead K562 cells depended on the number of NK cells and on the presence of IL-2. NK cells cytotoxic potential decreased gradually in the culture without IL-2. In contrast, NK cell-mediated cytotoxicity remained the same during the entire re-culture period after IL-2 re-application. CONCLUSION: Our study proved the efficacy of using cryopreserved ready-for-use NK cells in relapse treatment and the need for simultaneous administration of IL-2.

Postimplantation pressure testing and characterization of laser bonded glass/polyimide microjoints.

The stability of the laser bonded titanium coated glass/polyimide microjoints were studied in vivo by implanting on a rat brain surface for 10 days. In the current state, the strength of the joints were measured by a specially designed instrument called "pressure test" equipment where the samples were subjected to a variable pressure load (using high pressure nitrogen) controlled by a pressure regulator. The strength of the joints seems to degrade by about 28% as a result of soaking in rat brain. The bond degradation in rat brain implants is similar compared with those soaked in artificial cerebrospinal fluid (CSF) solution. Polyimide uptakes water through existing pores in it and also water gets in the joint region through the edges of the samples. Water might have caused oxidation of the chemical bonds which are thought to have formed by the laser fabrication process. A separate set of samples were created using same parameters for testing the hermeticity of the laser bonds. The samples were also exposed to rat brain CSF and were tested for hermiticity at the end of 10 days exposure time. It was observed that the implanted samples retained their hermeticity although the bond strength degraded by about 28%.

A neural cell culture study on thin film electrode materials.

Functional neural stimulation requires good interface between the neural cells and the electrode surfaces. In order to study the effect of electrode materials and surface structure on cell adhesion and biocompatibility, we cultured cortical neurons on thin films of platinum and iridium oxide. We used both flat, as-deposited and laser micro-structured films. The laser micro-structuring consisted of creating regular arrays of micro-bumps or holes with diameters of 4-5 mum and height of about 1.5 mum. The micro-bumps were fabricated onto platinum and iridium film surfaces deposited on borosilicate glass substrates, using mask-projection irradiation with single nano-second pulses from a KrF excimer laser (lambda = 248 nm). Amorphous and crystalline (deposited at 250 degrees C) IrO(2) films were deposited onto the laser micro-structured iridium films by pulsed-DC reactive sputtering to obtain micro-structured IrO(2) films. Cortical neurons isolated from rat embryo brain were cultured onto these film surfaces. Our results indicate that flat and micro-structured film surfaces are biocompatible and non-toxic for neural cell growth. The use of poly-D: -lysine as a mediator for cell adhesion onto the thin film surfaces is also discussed.