Development of a smart bioreactor for mammalian adherent cell expansion for application to cell therapy

开发用于哺乳动物贴壁细胞扩增的智能生物反应器以应用于细胞治疗

• 批准号: EP/H021442/1
• 负责人: Hua Ye
• 金额: $ 12.96万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2010
• 资助国家: 英国
• 起止时间: 2010 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=EP%2FH021442%2F1
• 关键词: Development; smart; bioreactor; mammalian; adherent

Cell therapy, particularly stem cell therapy, shows great promise for curing many diseases and repairing defective tissues in the human body. The problem currently is the difficulty in acquiring sufficient numbers of cells directly from a patient or donor. There is an urgent need to find a way to produce large quantities of high quality cells for cell therapy. The proposed research project will develop a smart bioreactor based on thermoresponsive polymer coated beads for culturing and expanding (increasing in cell number) adherent cells (as most cells, except blood cells, need to attach onto a surface to grow). The temperature sensitive polymer, poly(N-isopropylacrylamide) (PNIPAAm), has been found to be able to support cell adhesion and growth at 37 degree C, but when the temperature drops below 32 degree C, the cells will detach from the polymer and can thus be collected easily. This has distinctive advantages over the conventional cell culture methods in which adherent cells are harvested by destroying the proteins responsible for cell adhesion through the action of an enzyme such as trypsin. However, this enzyme also damages other crucial proteins on cell surfaces, which can affect cell growth and function. Work has been done to graft PNIPAAm to flat surfaces successfully and cells were able to grow into a thin sheet which was used to repair heart tissue defects in the human body.This project will look at how to graft PNIPAAm onto the surfaces of different types of small beads for cell culture. PNIPAAm grafted beads have already been used in chromatography for separating biological compounds, but very limited research has been done on using them for culturing human cells. Culturing adherent cells on small beads is routinely done both in research laboratories and in industry to expand cells, but the enzymatic treatment is needed to harvest cells at the end of cell culture.The thermosensitive beads to be developed in this project will enable separation of cells from beads without the damaging enzymatic treatment. In the project, different types of beads will be grafted with PNIPAAm and tested to see whether: (1) they have any toxic effects on cells, (2) they support cell growth on them, (3) cells can detach from the beads when the temperature drops below 32 degree C, and (4) cells still retain their function after being cultured on the beads. Also, a purpose-designed and fabricated bioreactor, a vessel to accommodate cells and beads and supply necessary nutrients and oxygen to the cells will be developed. This so-named smart bioreactor will be used to generate high quality cells suitable for cell therapy.

细胞疗法，特别是干细胞疗法，在治愈许多疾病和修复人体内有缺陷的组织方面显示出巨大的前景。目前的问题是，直接从患者或捐赠者那里获取足够数量的细胞是困难的。迫切需要找到一种方法来大量生产高质量的细胞用于细胞治疗。拟议的研究项目将开发一种基于热敏聚合物涂层小球的智能生物反应器，用于培养和扩大(细胞数量增加)贴壁细胞(因为大多数细胞，除血细胞外，需要附着在表面上才能生长)。温度敏感型聚合物聚N-异丙基丙烯酰胺(PNIPAAm)被发现能够在37摄氏度支持细胞黏附和生长，但当温度降至32摄氏度以下时，细胞将从聚合物中分离，因此易于收集。这比传统的细胞培养方法有明显的优势，在传统的细胞培养方法中，通过胰酶等酶的作用破坏负责细胞黏附的蛋白质来获得黏附细胞。然而，这种酶也会破坏细胞表面的其他关键蛋白质，从而影响细胞的生长和功能。已经成功地将PNIPAAm移植到平面上，细胞能够生长成一层薄片，用于修复人体心脏组织的缺陷。本项目将研究如何将PNIPAAm移植到不同类型的小球表面进行细胞培养。PNIPAAm接枝微珠已经用于层析分离生物化合物，但用于培养人类细胞的研究非常有限。在小球上培养贴壁细胞是研究实验室和工业中的常规做法，以扩大细胞，但在细胞培养结束时需要酶处理来获取细胞。本项目将开发的温敏小球将使细胞从小球中分离出来，而不需要破坏性的酶处理。在该项目中，将在不同类型的珠子上嫁接PNIPAAM并进行测试，以了解：(1)它们是否对细胞有任何毒性作用；(2)它们是否支持细胞在其上生长；(3)当温度降至32摄氏度以下时，细胞可以从珠子上分离；(4)细胞在珠子上培养后仍然保持其功能。此外，还将开发一种专门设计和制造的生物反应器，一种容纳细胞和珠子并为细胞提供必要营养和氧气的容器。这种所谓的智能生物反应器将被用来产生适合细胞治疗的高质量细胞。

项目成果

Bioreactor for stem cell culture using thermoresponsive microcarriers

使用热响应微载体进行干细胞培养的生物反应器

• 作者: Hua Ye (Co-Author)

Comprehensive Membrane Science and Engineering

• DOI: 10.1016/c2009-1-28385-7
• 发表时间: 2017-07
• 期刊: IEEE Access
• 影响因子: 3.9
• 作者: Hamed Karkhanechi

Fabrication of calcium phosphate fibres through electrospinning and sintering of hydroxyapatite nanoparticles

• DOI: 10.1016/j.matlet.2013.04.110
• 发表时间: 2013-09
• 期刊: Materials Letters
• 影响因子: 3
• 作者: P. Mouthuy;A. Crossley;H. Ye

Performances of a portable electrospinning apparatus.

• DOI: 10.1007/s10529-014-1760-6
• 发表时间: 2015-05
• 期刊: BIOTECHNOLOGY LETTERS
• 影响因子: 2.7
• 作者: Mouthuy, Pierre-Alexis;Groszkowski, Lukasz;Ye, Hua
• 通讯作者: Ye, Hua

Three-dimensional perfused tumour spheroid model for anti-cancer drug screening.

• DOI: 10.1007/s10529-016-2035-1
• 发表时间: 2016-08
• 期刊: Biotechnology letters
• 影响因子: 2.7
• 作者: Wan X;Li Z;Ye H;Cui Z
• 通讯作者: Cui Z