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碳酸化磷灰石表面的小鼠胚胎干细胞定殖

Mouse embryonic stem cell colonisation of carbonated apatite surfaces.

作者信息

Melville Amanda J, Harrison Janine, Gross Kārlis A, Forsythe John S, Trounson Alan O, Mollard Richard

机构信息

School of Physics and Materials Engineering, Monash University, Building 69, Clayton 3800, Melbourne, Australia.

出版信息

Biomaterials. 2006 Feb;27(4):615-22. doi: 10.1016/j.biomaterials.2005.06.028. Epub 2005 Aug 15.

Abstract

Apatites play a crucial role in the body and have been used extensively in biomedical implants. The influence on stem cell behaviour is not known and so this study will explore whether sintered carbonated apatites are favourable for propagation of stem cells. Different weight substitutions of carbonated apatite, specifically 2.5 wt% (2.5 wt%CAP) and 5 wt% (5 wt%CAP), were sintered and characterised prior to the investigation of their potential as a matrix for the support of mouse embryonic stem (ES) cells. Characterisation of the apatites included elemental analysis, X-ray diffraction, surface roughness, specific surface area, density, and solubility. The ability of carbonated apatite to support mouse ES cell colonisation and maintenance in the presence of leukaemia inhibitory factor was determined by an enumeration of live versus dead cells within a population, and immunoreactivity to Oct4, a transcription factor and stem cell marker, following growth on each matrix. It was found that while both compositions allowed for the colonisation of mouse ES cells, the cells were not maintained in an undifferentiated state, as evidenced by a reduction in the number of cells staining positive for Oct4 expression. This study shows that an increase in carbonate content within sintered apatites leads to a higher cell number, a desired aspect for stem cells to populate scaffolds intended for tissue engineering. This study presents carbonated apatites as a suitable matrix for the initial colonisation and differentiation of ES cells for tissue engineering applications.

摘要

磷灰石在人体中起着至关重要的作用,并且已广泛应用于生物医学植入物。其对干细胞行为的影响尚不清楚,因此本研究将探讨烧结碳酸化磷灰石是否有利于干细胞的增殖。在研究其作为支持小鼠胚胎干细胞(ES细胞)的基质的潜力之前,对碳酸化磷灰石进行了不同重量百分比的替代烧结,具体为2.5重量%(2.5 wt%CAP)和5重量%(5 wt%CAP),并对其进行了表征。磷灰石的表征包括元素分析、X射线衍射、表面粗糙度、比表面积、密度和溶解度。通过对群体中活细胞与死细胞的计数,以及在每种基质上生长后对转录因子和干细胞标志物Oct4的免疫反应性,确定了碳酸化磷灰石在白血病抑制因子存在下支持小鼠ES细胞定植和维持的能力。结果发现,虽然两种成分都允许小鼠ES细胞定植,但细胞并未维持在未分化状态,Oct4表达阳性染色细胞数量减少证明了这一点。本研究表明,烧结磷灰石中碳酸盐含量的增加会导致细胞数量增加,这是干细胞在组织工程支架中增殖的一个理想方面。本研究表明,碳酸化磷灰石是用于组织工程应用的ES细胞初始定植和分化的合适基质。

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