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

Year 2018, Volume: 4 Issue: 3, 145 - 151, 04.07.2018

Ozan Karaman , Ziyşan Buse Yaralı

https://doi.org/10.18621/eurj.342109

Cited By: 3

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 7^th day after micro-tissue
formation, live/dead cells analysis was conducted. Micrograph taken on days 1,
3, 5 and 7^th 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.

Keywords

micro-tissue, three-dimensional cell culture, serum concentration

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