Impact of long term propagation in 3D culture on the phenotype of existing human cell lines
3D 培养中的长期繁殖对现有人类细胞系表型的影响
基本信息
- 批准号:1649096
- 负责人:
- 金额:--
- 依托单位:
- 依托单位国家:英国
- 项目类别:Studentship
- 财政年份:2015
- 资助国家:英国
- 起止时间:2015 至 无数据
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
Tissue-specific architecture, mechanical and biochemical cues and cell-cell interactions are altered or lost under the simplified conditions of two-dimensional (2D) cell culture. Three-dimensional (3D) cell cultures attempt to mimic the in vivo situation by preserving the 3D integrity of individual cells and enabling cells to create their own niche in conjunction with the extracellular matrix. There is good evidence to show that 3D models can be used to study physiology where tissue-like function can be maintained. It is common practice however to expand populations of cells in 2D culture initially (particularly cell lines) prior to seeding such cells into 3D culture models. In such circumstances, cells are simultaneously: a) adapting to their new surroundings (2D to 3D growth transition); and b) being challenged and/or examined as part of the 3D cell culture assay (for example, a drug cytotoxicity test). This makes it difficult to decipher the precise outcome of the assay given that cells are concurrently exposed to two such major variables.We hypothesise that cell populations initially propagated and expanded within a 3D environment are more likely to adapt to their surroundings when seeded into a 3D culture model or transplanted in vivo (i.e. without the need for 2D to 3D transition). In this project we will collaborate with ECACC Culture Collections, an organization that supplies cell lines to researchers. The propagation of popular cell lines that are grown routinely as adherent 2D monolayers (e.g. A549 lung epithelium; MCF7 breast tumour; both established and widely understood), will be maintained in a novel system that enables continual cell passaging in 3D to convert their phenotype to a more native 3D state. Specifically, the student will: 1) (0-18 mths) optimize methods to continually passage A549 and MCF7 human cell lines in 3D culture; 2) (6-24 mths) Characterize the cell phenotype by detailed examination of cell morphology, cytoskeletal structure, expression of integrin/caherins and focal adhesion proteins, expression of markers associated with 3D phenotype known to be lost in 2D culture; 3) (18-30 mths) conduct a 3-way comparison of 2D vs 3D vs in vivo (e.g. MCF7 tumour tissue) examining key structural cell proteins known to be differentially regulated in 2D and 3D culture; 4) (24-36mths) test the ability of the 2D vs 3D propagated cells to produce 3D structures when grown in alternative technologies designed to support 3D cell growth in vitro, e.g. scaffolds, and spheroid cultures; 5) (30-36 mths) Assess the ability of 2D & 3D propagated cells to form xenograft tumours when transplanted into immune deficient mice. The growth rate, size and tissue composition will be compared. The project will provide new information of how the growth environment plays an important role in the basic biology of cultured cells. The student will gain knowledge of cell science and training in cellular and molecular techniques with academic and industrial partners. It is anticipated that these new approaches for maintaining cells will be immediately beneficial to biologists looking to improve their in vitro model.
在二维(2D)细胞培养的简化条件下,组织特异性结构,机械和生化线索以及细胞-细胞相互作用被改变或丢失。三维(3D)细胞培养试图通过保持单个细胞的三维完整性,并使细胞能够与细胞外基质一起创造自己的生态位来模拟体内情况。有充分的证据表明,3D模型可以用于研究生理学,其中可以维持组织样功能。然而,在将细胞播种到3D培养模型之前,通常的做法是首先在2D培养中扩增细胞群(特别是细胞系)。在这种情况下,细胞同时:a)适应新的环境(2D到3D的生长过渡);以及b)作为3D细胞培养试验(例如,药物细胞毒性试验)的一部分进行挑战和/或检查。鉴于细胞同时暴露于两个这样的主要变量,这使得破译测定的精确结果变得困难。我们假设最初在3D环境中繁殖和扩增的细胞群在播种到3D培养模型或在体内移植(即不需要2D到3D转换)时更有可能适应周围环境。在这个项目中,我们将与ECACC Culture Collections合作,这是一个向研究人员提供细胞系的组织。通常作为粘附二维单层生长的常见细胞系(例如A549肺上皮;MCF7乳腺肿瘤;两者都已建立并被广泛理解)的繁殖将在一个新的系统中维持,该系统使细胞在3D中连续传代,将其表型转化为更天然的3D状态。具体而言,学生将:1)(0-18个月)优化方法,在3D培养中连续传代A549和MCF7人类细胞系;2)(6-24个月)通过详细检查细胞形态、细胞骨架结构、整合素/钙粘着素和局灶黏附蛋白的表达、已知在2D培养中丢失的与3D表型相关的标志物的表达来表征细胞表型;3)(18-30个月)进行二维、三维和体内(如MCF7肿瘤组织)的三向比较,检查已知在二维和三维培养中差异调节的关键结构细胞蛋白;4)(24-36个月)测试2D与3D繁殖细胞在用于支持体外3D细胞生长的替代技术中生长时产生3D结构的能力,例如支架和球形培养;5)(30-36个月)评估2D和3D增殖细胞移植到免疫缺陷小鼠体内形成异种移植物肿瘤的能力。比较生长速度、大小和组织组成。该项目将提供关于生长环境如何在培养细胞的基础生物学中发挥重要作用的新信息。学生将获得细胞科学知识,并与学术和工业合作伙伴一起接受细胞和分子技术方面的培训。可以预期,这些维持细胞的新方法将立即有利于生物学家改善他们的体外模型。
项目成果
期刊论文数量(0)
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科研奖励数量(0)
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专利数量(0)
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其他文献
吉治仁志 他: "トランスジェニックマウスによるTIMP-1の線維化促進機序"最新医学. 55. 1781-1787 (2000)
Hitoshi Yoshiji 等:“转基因小鼠中 TIMP-1 的促纤维化机制”现代医学 55. 1781-1787 (2000)。
- DOI:
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- 影响因子:0
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LiDAR Implementations for Autonomous Vehicle Applications
- DOI:
- 发表时间:
2021 - 期刊:
- 影响因子:0
- 作者:
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吉治仁志 他: "イラスト医学&サイエンスシリーズ血管の分子医学"羊土社(渋谷正史編). 125 (2000)
Hitoshi Yoshiji 等人:“血管医学与科学系列分子医学图解”Yodosha(涉谷正志编辑)125(2000)。
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Effect of manidipine hydrochloride,a calcium antagonist,on isoproterenol-induced left ventricular hypertrophy: "Yoshiyama,M.,Takeuchi,K.,Kim,S.,Hanatani,A.,Omura,T.,Toda,I.,Akioka,K.,Teragaki,M.,Iwao,H.and Yoshikawa,J." Jpn Circ J. 62(1). 47-52 (1998)
钙拮抗剂盐酸马尼地平对异丙肾上腺素引起的左心室肥厚的影响:“Yoshiyama,M.,Takeuchi,K.,Kim,S.,Hanatani,A.,Omura,T.,Toda,I.,Akioka,
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