RESUMO
The study focused on activation of lymphocytes derived from healthy donors and cancer patients in the medium supplemented with IL-2 and IL-15 at low concentrations. We studied morphological features of cultured cells, their viability, and proliferative and functional activities during culturing for up to 14 days. In comparison with lymphocytes derived from cancer patients, white blood cells derived from healthy donors demonstrated a number of advantages under the given culturing conditions such as higher viability, greater proliferative activity, and the potency for earlier activation; moreover, during activation they secreted the anti-inflammatory cytokines IL-4 and IL-10 at greater concentrations.
Assuntos
Interleucina-15/farmacologia , Interleucina-2/farmacologia , Linfócitos/efeitos dos fármacos , Células Cultivadas , Humanos , Imunoterapia , Transdução de Sinais/efeitos dos fármacosRESUMO
Microorifice-based fusion makes use of electric field constriction to assure high-yield one-to-one fusion of selected cell pairs. The aim of this paper is to verify feasibility of high-yield cell fusion on a microfluidic chip. This paper also examines viability of the fusant created on the chip. We fabricated a microfluidic chip to fuse selected cell pairs and to study postfusion behavior. We used a self-forming meniscus-based fabrication process to create microorifice with a diameter of 2-10 microm on the vertical walls in a microfluidic channel. When 1 MHz was applied to electrodes located on both sides of the microorifice, dielectrophoretic force attracted the cells toward microorifice to form a cell pair. Once the cells get into contact, fusion pulse was applied. Real time imaging of cells during fusion and cytoplasmic dye transfer between cells indicated success of cell fusion. We found that when high frequency voltage for dielectrophoresis was swept from 1 MHz to 10 kHz in 100 micros, cell fusion was initiated. The effective electric field strength was 0.1-0.2 kV/cm. We analyzed viability by imaging fusant going into cell division phase after 48 h of incubation. We conclude that fabricated microfluidic chip is suitable for high-yield one-to-one fusion and creation of viable fusants. This technology should be a useful tool to study fusion phenomena and viability of fusants, as it allows imaging of the cells during and after the fusion.
Assuntos
Fusão Celular/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Animais , Engenharia Biomédica , Fusão Celular/métodos , Linhagem Celular , Sobrevivência Celular , Eletricidade , Desenho de Equipamento , Humanos , Células Jurkat , Camundongos , Técnicas Analíticas Microfluídicas/métodos , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Microscopia de Contraste de FaseRESUMO
RNA interference (RNAi) is among the most particular mechanisms of gene expression regulation. Besides, small interfering RNAs are significant players in cell defence either from viral infection or retrotransposons. Medical utilization of RNAi gives a handful of ways to cure viral and oncological illnesses. RNA interference, also, represents a useful tool for research, because it allows quick production of monogene functional knockouts. In this review we describe the most recent conceptions about RNAi mechanisms and actual approaches for it's usage.
Assuntos
Neoplasias/genética , Interferência de RNA , RNA Interferente Pequeno/genética , Retroelementos/genética , Viroses/genética , Animais , Biotecnologia , Humanos , Neoplasias/terapia , Viroses/terapiaRESUMO
This study reports on a novel method to improve the strength of apatitic bone cements. The liquid phase of Biocement-H was modified with commercial superplasticizers. The results showed that small additions, i.e. 0.5 vol%, in the aqueous liquid phase improved the maximum compressive strength of Biocement-H (35 MPa) by 71%, i.e. 60 MPa. Moreover, the addition of high amounts of superplasticizers, i.e. 50 vol.%, allowed for a significant reduction of the liquid-to-powder ratio from 0.32 to 0.256 mL/g, without affecting the maximum strength and/or the workability of the cement. These results open up new ways to develop injectable and high-strength apatitic bone cements for load-bearing applications.