Larsen Benjamin Endré, Pettersen Erik Olai, Tønnesen Hanne Hjorth, Melvik Jan Egil
School of Pharmacy, University of Oslo, Oslo, Norway.
Cell Prolif. 2015 Jun;48(3):318-29. doi: 10.1111/cpr.12178. Epub 2015 Mar 23.
The main objective of this paper was to investigate whether the oxygen consumption rate (OCR) of cells entrapped in alginate hydrogels depends on presence of soluble factors present in foetal bovine serum (FBS).
Pericellular oxygen concentrations were measured using a photochemical oxygen sensor inserted into bioconstructs made from different formulations of alginate, containing T-47D cells. Cell count was corrected for viability as determined by cell uptake and exclusion of standard live/dead fluorophores, in sections of freshly prepared biostructures. Based on concentration data, OCR of the embedded cells was calculated according to a simple algorithm.
OCR was found to vary significantly between the different formulations investigated. Inclusion of high concentrations of FBS in the biostructure matrices elicited significantly higher OCRs, in guluronate-rich gels similar to those previously found in monolayer culture. Guluronate-rich gels also generally permitted highest OCR. Respiration also had a falling tendency with increasing alginate concentration and elastic modulus.
Presence of FBS in biostructure matrices elicited higher OCR in T-47D cells. Formulation of biostructures must consider differential diffusion of macromolecular substances.
本文的主要目的是研究包裹在藻酸盐水凝胶中的细胞的耗氧率(OCR)是否取决于胎牛血清(FBS)中可溶性因子的存在。
使用插入由不同配方藻酸盐制成的生物构建体(含有T-47D细胞)中的光化学氧传感器测量细胞周围的氧浓度。在新鲜制备的生物结构切片中,根据细胞摄取和标准活/死荧光团的排除情况对细胞活力进行校正后计算细胞数量。根据浓度数据,按照简单算法计算包埋细胞的OCR。
在所研究的不同配方之间,OCR存在显著差异。在生物结构基质中加入高浓度FBS会引发显著更高的OCR,在富含古洛糖醛酸的凝胶中,其OCR与先前在单层培养中发现的相似。富含古洛糖醛酸的凝胶通常也允许最高的OCR。随着藻酸盐浓度和弹性模量的增加,呼吸也有下降趋势。
生物结构基质中FBS的存在会引发T-47D细胞中更高的OCR。生物结构的配方必须考虑大分子物质的差异扩散。