COMPOSITION AND FUNCTIONS OF EXTRACELLULAR PROTEINS IN LEAF APOPLAST AND IN VITRO CELL CULTURE MEDIUM OF TARTARY BUCKWHEAT

Abstract Stem cell factor is one of the most important growth factors for human hematopoietic stem cells (HSC). Recombinant human stem cell factor (rhSCF) can stimulate HSC expansion and regeneration in vitro, when it is used in combination with other cytokines like Flt-3L and TPO. However, the specific structural region(s) of the rhSCF protein that are critical for its function in HSC expansion are still unknown. Few studies have addressed this problem, to date. We have recently reported the production of a novel monoclonal antibody (named 23C8) against rhSCF, and the demonstration that 23C8 could inhibit the ability of rhSCF to enhance HSC expansion. Here, we report the identification of a short polypeptide from rhSCF that contains the epitope for binding to 23C8, and, like the full-length rhSCF, is able to stimulate the expansion of umbilical cord blood (UCB)-derived CD34+ cells. Twelve short polypeptides were designed and synthesized, which cover the full length of rhSCF, with 3-5 amino acids overlaps. 23C8 was collected from hybridoma cell culture medium and further purified using protein G affinity chromatography. ELISA was used to identify the polypeptide(s) that positively react with 23C8 among all the synthesized polypeptides. In addition, the effects of the synthetic polypeptides on human HSC expansion capacity were evaluated by supplementing the cell culture medium with 100 ng/ml of a given polypeptide. Total cell number and CD34+ cell number of each group were monitored on day 6. Our novel anti-SCF monoclonal antibody (23C8) partially blocked SCF’s function in human UCB CD34+ cell expansion. Of all the polypeptides analyzed, only one, named P0, corresponding to the SCF protein sequence at residues 40 to 57, was recognized by 23C8 during ELISA. P0, like the full-length rhSCF, enhanced expansion of CD34+ cells derived from human UCB. P0 addition increased the numbers of total nucleated cells and CD34+ cells by 10.58±0.86 and 4.63±0.43 folds, respectively. For comparison, the extents of increases in cell numbers in the vehicle control group was 3.15±0.99 fold (total nucleated cells) and 1.07±0.11 fold (CD34+ cells), respectively. Residues 40-57 of hrSCF comprise a critical functional region for its ability to enhance expansion of human UCB CD34+ cells in vitro. The short P0 peptide is a potential candidate for development as a synthetic substitute for rhSCF in clinic applications. Disclosures Jiang: Biopharmagen.corp: Employment. Jiang:Biopharmagen.corp: Employment.

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Exposure Setup and Dosimetry for a Study on Effects of Mobile Communication Signals on Human Hematopoietic Stem Cells in vitro

Advances in Radio Science ◽

10.5194/ars-15-207-2017 ◽

2017 ◽

Vol 15 ◽

pp. 207-213

Author(s):

Martina Rohland ◽

Kai Baaske ◽

Katharina Gläser ◽

Henning Hintzsche ◽

Helga Stopper ◽

...

Keyword(s):

Stem Cells ◽

Cell Culture ◽

Hematopoietic Stem Cells ◽

Mobile Communication ◽

Culture Medium ◽

Cell Culture Medium ◽

Scattering Parameter ◽

Hematopoietic Stem ◽

Communication Signals

Abstract. In this paper we describe the design of an exposure setup used to study possible non-thermal effects due to the exposure of human hematopoietic stem cells to GSM, UMTS and LTE mobile communication signals. The experiments are performed under fully blinded conditions in a TEM waveguide located inside an incubator to achieve defined environmental conditions as required for the living cells. Chamber slides containing the cells in culture medium are placed on the septum of the waveguide. The environmental and exposure parameters such as signal power, temperatures, relative humidity and CO2 content of the surrounding atmosphere are monitored permanently during the exposure experiment. The power of the exposure signals required to achieve specific absorption rates of 0.5, 1, 2 and 4 W kg−1 are determined by numerical calculation of the field distribution inside the cell culture medium at 900 MHz (GSM), 1950 MHz (UMTS) and 2535 MHz (LTE). The dosimetry is verified both with scattering parameter measurements on the waveguide with and without containers filled with cell culture medium and with temperature measurements with non-metallic probes in separate heating experiments.

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Investigation of quercetin stability in cell culture medium: Role in in vitro experiment

African Journal of Pharmacy and Pharmacology ◽

10.5897/ajpp11.721 ◽

2012 ◽

Vol 6 (14) ◽

Author(s):

Jun Hu

Keyword(s):

Cell Culture ◽

Culture Medium ◽

Cell Culture Medium ◽

Vitro Experiment ◽

In Vitro Experiment

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Characterisation of NiTi Orthodontic Archwires Surface after the Simulation of Mechanical Loading in CACO2-2 Cell Culture

Coatings ◽

10.3390/coatings9070440 ◽

2019 ◽

Vol 9 (7) ◽

pp. 440 ◽

Cited By ~ 2

Author(s):

Nikola Lepojević ◽

Ivana Šćepan ◽

Branislav Glišić ◽

Monika Jenko ◽

Matjaž Godec ◽

...

Keyword(s):

Cell Culture ◽

Mechanical Loading ◽

Inductively Coupled Plasma ◽

Culture Medium ◽

Cell Culture Medium ◽

Nickel Ions ◽

Inductively Coupled ◽

Icp Ms ◽

Orthodontic Archwires

Nickel-titanium (NiTi) orthodontic archwires are crucial in the initial stages of orthodontic therapy when the movement of teeth and deflection of the archwire are the largest. Their great mechanical properties come with their main disadvantage—the leakage of nickel. Various in vitro studies measured nickel leakage from archwires that were only immersed in the medium with little or minimal simulation of all stress and deflection forces that affect them. This study aims to overcome that by simulating deflection forces that those archwires are exposed to inside the mouth of a patient. NiTi orthodontic archwires were immersed in CACO2-2 cell culture medium and then immediately loaded while using a simulator of multiaxial stress for 24 h. After the experiment, the surface of the NiTi orthodontic archwires were analysed while using scanning electron microscopy (SEM) and auger electron spectroscopy (AES). The observations showed significant microstructural and compositional changes within the first 51 nm thickness of the archwire surface. Furthermore, the released nickel and titanium concentrations in the CACO2-2 cell culture medium were measured while using Inductively Coupled Plasma Mass Spectroscopy (ICP-MS). It was found out that the level of released nickel ions was 1.310 µg/L, which can be assigned as statistically significant results. These data represent the first mention of the already detectable release of Ni ions after 24 h during the simulation of mechanical loading in the CACO2-2 cell culture medium, which is important for clinical orthodontic praxis.

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Using 2H2O to study the influence of feeding on protein synthesis: effect of isotope equilibration in vivo vs. in cell culture

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00580.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. E1277-E1283 ◽

Cited By ~ 59

Author(s):

Danielle A. Dufner ◽

Ilya R. Bederman ◽

Daniel Z. Brunengraber ◽

Nadia Rachdaoui ◽

Faramarz Ismail-Beigi ◽

...

Keyword(s):

Cell Culture ◽

Protein Synthesis ◽

Steady State ◽

Culture Medium ◽

Body Water ◽

Cell Culture Medium ◽

Plasma Albumin ◽

Single Meal

We previously reported that 2H2O can be used to measure rates of protein synthesis during prolonged steady-state conditions (Previs SF, Fatica R, Chandramouli V, Alexander JC, Brunengraber H, and Landau BR. Am J Physiol Endocrinol Metab 286: E665-E672, 2004). The underlying premise of our method is that following the administration of 2H2O, 2H atoms in body water rapidly equilibrate with free alanine before it is incorporated into newly synthesized proteins. We have now directly examined whether 2H2O can be used to measure the influence of a single meal on protein synthesis. In addition, we have compared the use of 2H2O for measuring rates of protein synthesis in vivo vs. in cell culture. Using a rat model, we observed rapid equilibration between 2H in body water and free alanine; therefore we were able to study the response of protein synthesis to a single meal. We observed that ∼50% of the plasma albumin that is synthesized over the course of 24 h is made within ∼5 h after eating (in rats trained to eat a complete 24-h ration of food in a single meal). Contrary to what we observed in vivo, feeding (the replenishment of cell culture medium) does influence the use of 2H2O for in vitro studies. In particular, since there can be slow equilibration of 2H between water and alanine in the cell culture medium, special consideration must be made to avoid underestimating the rate of protein synthesis in vitro.

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The study of biological activity nanomaterial in cultivation conditions pigs sperm and oocytes in vitro

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v20.775 ◽

1970 ◽

pp. 256-260 ◽

Cited By ~ 2

Author(s):

O. V. Shcherbak ◽

A. B. Ziuziun ◽

O. S. Osypchuk ◽

S. I. Kovtun ◽

N. P. Halahan ◽

...

Keyword(s):

Cell Culture ◽

Biological Activity ◽

Culture Medium ◽

Culture Conditions ◽

Cell Culture Medium ◽

Animal Genetic Resources ◽

Cultivation In Vitro ◽

Breeding And Genetics ◽

Positive Effect

Aim. To assess the biological activity of nanomaterial UFS/BSA/NANA of culture conditions with different concentrations of spermatozoa and the oocyte-cumulus complexes of pigs in vitro. Methods. For study biological activity in experiments used ejaculate cryopreserved sperms of three boars (Bank Animal Genetic Resources Institute of Animal Breeding and Genetics nd. a. M.V. Zubets of National Academy of Agrarian Science of Ukraine) and freshly prepared oocytes pigs. Results. In study effect of different concentrations NM (UFS/BSA/NANA) on viability gametes boar. The most active sperms when we add UFS/BSA/NANA in concentration 0.001 %. Adding UFS/BSA/NANA 0.1 % concentration in the cell culture medium has provided formation mature pigs oocytes for significantly higher level, which was 20.2 % more compared to the control and 18.7 % higher, when we add UFS/BSA/NANA in concentration 0.001 %. Conclusions. It is demonstrated the positive effect on defrosted boars sperms with adding UFS/BSA/NANA in concentration 0.001 %, their biological activity increased by 13.3 % compared with control. It is demonstration that add UFS/BSA/NANA in concentration 0.1 % in cell culture medium with pig’s oocytes, level of maturation increased by 20.2 %.Keywords: oocytes, sperms, pigs, cultivation in vitro, ultra-fine silica (UFS).

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