scholarly journals Preconditioning of Bioactive Glasses before Introduction to Static Cell Culture: What Is Really Necessary?

2020 ◽  
Vol 3 (2) ◽  
pp. 38 ◽  
Author(s):  
Frederike Hohenbild ◽  
Marcela Arango-Ospina ◽  
Arash Moghaddam ◽  
Aldo R. Boccaccini ◽  
Fabian Westhauser
Keyword(s):  
Cell Culture ◽  
Stromal Cells ◽  
Culture Medium ◽  
Cell Culture Medium ◽  
Bioactive Glasses ◽  
Experimental Protocols ◽  
Passivation Time ◽  
Vitro Analysis ◽  
In Vitro Analysis

Due to their high bioreactivity, the in-vitro analysis of bioactive glasses (BGs) can be challenging when it comes to maintaining a physiological pH. To improve BG biocompatibility, a heterogenic spectrum of preconditioning approaches, such as “passivation” of the BGs by incubation in cell culture medium, are used but have never been directly compared. In this study, the effect of passivation periods of up to 72 h on pH alkalization and viability of human bone marrow-derived mesenchymal stromal cells was evaluated to determine a time-efficient passivation protocol using granules based on the 45S5-BG composition (in wt%: 45.0 SiO2, 24.5 Na2O, 24.5 CaO, 6.0 P2O5) in different concentrations. pH alkalization was most reduced after passivation of 24 h. Cell viability continuously improved with increasing passivation time being significantly higher after passivation of at least 24 h compared to non-passivated 45S5-BG and the necessary passivation time increased with increasing BG concentrations. In this setting, a passivation period of 24 h presented as an effective approach to provide a biocompatible cell culture setting. In conclusion, before introduction of BGs in cell culture, different passivation periods should be evaluated in order to meet the respective experimental settings, e.g., by following the experimental protocols used in this study.

Fluoride-containing bioactive glasses and Bioglass® 45S5 form apatite in low pH cell culture medium

2014 ◽  
Vol 119 ◽  
pp. 96-99 ◽  
Author(s):  
Furqan A. Shah ◽  
Delia S. Brauer ◽  
Nikita Desai ◽  
Robert G. Hill ◽  
Karin A. Hing
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Low Ph ◽  
Cell Culture Medium ◽  
Bioactive Glasses

scholarly journals Silk fibroin induces chondrogenic differentiation of canine adipose–derived multipotent mesenchymal stromal cells/mesenchymal stem cells

2019 ◽  
Vol 10 ◽  
pp. 204173141983505 ◽  
Author(s):  
Metka Voga ◽  
Natasa Drnovsek ◽  
Sasa Novak ◽  
Gregor Majdic
Keyword(s):  
Stem Cells ◽  
Cell Culture ◽  
Stromal Cells ◽  
Chondrogenic Differentiation ◽  
Culture Medium ◽  
Collagen Type ◽  
Multipotent Mesenchymal Stromal Cells ◽  
Culture Conditions ◽  
Standard Cell ◽  
Cell Culture Medium

Under appropriate culture conditions, mesenchymal stem cells (MSC), also called more properly multipotent mesenchymal stromal cells (MMSC), can be induced toward differentiation into different cell lineages. In order to guide stem cell fate within an environment resembling the stem cell niche, different biomaterials are being developed. In the present study, we used silk fibroin (SF) as a biomaterial supporting the growth of MMSC and studied its effect on chondrogenesis of canine adipose–derived MMSC (cADMMSC). Adipose tissue was collected from nine privately owned dogs. MMSC were cultured on SF films and SF scaffolds in a standard cell culture medium. Cell morphology was evaluated by scanning electron microscopy (SEM). Chondrogenic differentiation was evaluated by alcian blue staining and mRNA expression of collagen type 1, collagen type 2, Sox9, and Aggrecan genes. cADMMSC cultured on SF films and SF scaffolds stained positive using alcian blue. SEM images revealed nodule-like structures with matrix vesicles and fibers resembling chondrogenic nodules. Gene expression of chondrogenic markers Sox9 and Aggrecan were statistically significantly upregulated in cADMMSC cultured on SF films in comparison to negative control cADMMSC. This result suggests that chondrogenesis of cADMMSC could occur when cells were grown on SF films in a standard cell culture medium without specific culture conditions, which were previously considered necessary for induction of chondrogenic differentiation.

A Structure-Function Study of the Activity of Stem Cell Factor in Stimulating the Expansion of Cord Blood CD34+ Cells

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 3501-3501
Author(s):  
Bin Shen ◽  
Wenhong Jiang ◽  
Jie Fan ◽  
Wei Dai ◽  
Xinxin Ding ◽  
...  
Keyword(s):  
Cell Culture ◽  
Monoclonal Antibody ◽  
Stem Cell ◽  
Stem Cell Factor ◽  
Culture Medium ◽  
Cell Number ◽  
Full Length ◽  
Cell Culture Medium ◽  
Cd34 Cells

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.

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

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.

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

2020 ◽  
Author(s):  
Н.И. Румянцева ◽  
A.И. Валиева ◽  
A.Н. Акулов ◽  
A.В. Лайков ◽  
Ю.A. Костюкова ◽  
...  
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Extracellular Proteins ◽  
Tartary Buckwheat ◽  
Cell Culture Medium ◽  
In Vitro Cell Culture ◽  
Leaf Apoplast
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