Determination of minimum serum concentration to develop scaffold free micro-tissue

Abstract

Objective. Formation of three-dimensional (3D) micro-tissues without scaffolds are widely used not only to define in vivo tissue formation mechanisms but also the development of different tissue-specific drugs. However, depending on high serum and growth factor concentrations, it would be hard to identify major effective biological cues on micro-tissue formation. The aim of the study is to determine the effect of different serum concentrations on Human Umbilicial Vein Endothelial Cells (HUVECs) micro-tissue formation. Methods. Micro-tissue of HUVEC line was formed by using 3D petri dish technique with medium containing 0%, 1%, 5% and 10% fetal bovine serum (FBS). On the 7th day after micro-tissue formation, live/dead cells analysis was conducted. Micrograph taken on days 1, 3, 5 and 7th of micro-tissue formation were determined by image analysis with ImageJ. Results. Sizes of micro-tissue formed with 0% FBS on day 1 and 3 determined as 277 ± 12 µm and 279 ± 20 µm, respectively; however, especially on day 7 micro-tissue size significantly decreased to 229 ± 6 µm. When live/dead analysis results were examined, high cell viability was observed in 5% and 10% FBS concentration. Although micro-tissue like structures were observed in 0% and 1% FBS concentrations dead cell ratio considerably increased compared to 5% and 10% FBS concentration. Conclusions. It has been determined that 0% and 1% serum are appropriate for determining the efficacy of biomimetic peptides and different extracellular matrix proteins on micro-tissue formation parameters of HUVEC. High cell viability in micro-tissues was observed with 5% and 10% serum concentrations.

Regulatory effects of laminin derived peptide on microtissue formation for tissue engineered scaffold-free constructs

“…At the end of 7th day, less efficacy was seen in 3 mM YIGSR peptide concentration compared with 1.5 mM peptide concentration. Studies of Martin et al [26], Karaman and Yarali [27], and Yarali et al [28] demonstrated that formation of SFMs develops depending on the diameter enlargement of SFMs. In light of that information, we concluded that both of YIGSR peptide concentrations provide enlargement of HUVEC SFMs and contributed to developing SFMs formation until the end of the 4th day.…”

“…Martin, F. et al showed that the differentiation state and development of the generated SFMs can be identified by diameter calculation and SFMs maturation [26]. Moreover, Karaman and Yarali [27] demonstrated that SFMs diameter measurement can relate to SFMs growth and viability analysis. Therefore, diameter analysis can be used to identify healthy growing SFMs [27,28].…”

“…Moreover, Karaman and Yarali [27] demonstrated that SFMs diameter measurement can relate to SFMs growth and viability analysis. Therefore, diameter analysis can be used to identify healthy growing SFMs [27,28]. In this context, the diameter and viability analyzes have been performed on HUVEC SFMs with different YIGSR peptide concentrations.…”

“…To understand the effects of different YIGSR concentrations to the viability of HUVEC SFMs, viability assay was performed. The protocol was applied as described in previous studies [27,28]. The rate of live or dead cells in microtissue was indicated by Double Staining Kit (Dojindo, Molecular Technologies, Inc, Japan).…”

Biodegradable and Biocompatible Graphene-based Scaffolds for Functional Neural Tissue Engineering: A Strategy Approach Using Dental Pulp Stem Cells and Biomaterials

“…This could be due to the formation of protein corona on the GO surface which in turn influence the toxicity of GObased materials [61,62]. Furthermore, studies have indicated that serum proteins can have an interfering effect on interactions between nanoparticles, cells and biological molecules [63][64][65]. Accordingly, Lesniak et al [64] demonstrated that various protein corona formed on silica nanoparticles modified cell adhesion, cellular uptake, and toxicity, which is determined by the serum protein concentration.…”