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Minusheet Perfusion Culture System -细胞3D培养灌注系统,3D灌流灌注共培养系统，灌流梯度培养系统

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系统介绍 
Minusheet三维灌流系统设计有专门的封闭式的培养小室，联合蠕动泵，采用灌流系统进行体内模拟细胞培养，

使细胞能够聚集生长，形成组织样结构，很好的模拟体内生理状况，可以更直观的反映细胞生物学和功能，

更准确的构建靶组织模型，不仅可以培养单一细胞，而且还可以进行多种细胞的共培养。 
利用3D细胞培养技术使细胞形成3D微组织，从而更好地模拟体内细胞生长情况，更直观的反映细胞生物学和功能，

更准确的构建靶组织模型。 
可用于几乎任何类型细胞的3D培养，包括细胞系、原代细胞、干细胞及多种细胞共培养。应用范围涉及癌症研究及

药物分析，细胞生物学研究和组织工程，吸收、分布、新陈代谢、排泄及毒性测试（ADME-TOX)，药物代谢动力学（DMPK）。
传统的细胞培养瓶/皿依赖于培养箱提供CO2和湿度环境进行生长，而这些条件由于没有得到严格的控制，使得模拟的生

长环境完不同于严格意义上的体内环境，从而导致实验数据存在一定的偏差；且传统培养容器的操作和培养的开放性，

不能及时保持培养液的新鲜、不能保证代谢物的及时排除。容易导致细胞发生污染，而3D（三维）细胞培养克服了这些问题,

该系统自带的热板和气体交换模块，实时保证了细胞所需要的温湿度,适合细胞在保持养分不变的长期培养，

显微腔室便于实时培养观察，梯度培养腔室使得培养环境更类体内环境。

系统亮点

1. 充分的养分保障及时代谢物的排除，提高了细胞密度

连续灌注的培养基提供了充分的营养成分, 并可带走代谢产物, 同时, 细胞保留在反应器系统中,可以达到很高的细胞密度

1. 不用CO2培养箱提供温湿度环境，该系统自带的热板和气体交换模块，实时保证了细胞所需要的温湿度

1. 应用细胞类型广泛，几乎可以应用到任何类型细胞的3D培养。

图1. 细胞类型

3)可以联用显微镜 
Chamber上下两侧都是透明的，可以联合显微镜直接观察，甚至在灌流的过程中也可以观察。

图2. Chambers类型大

4).可以采用梯度培养，不同种类的细胞共培养。 
可以采用不同的培养基对细胞进行灌冲，这样能更好的模拟体内的环境，实验更准确。

图3.梯度灌流

图4. 两种细胞的共培养
4).可以长期培养 
样品可以灌注培养高达数月，可以进行周期较长的实验。
应用范围 
(1).可用于几乎任何类型细胞的3D培养，包括细胞系、原代细胞、干细胞及多种细胞共 
培养。 
(2). 癌症研究及药物分析
(3). 细胞生物学研究和组织工程
(4). 吸收、分布、新陈代谢、排泄及毒性测试（ADME-TOX)
(5). 药物代谢动力学（DMPK
Tannic acid label indicates abnormal cell development coinciding with regeneration 
of renal tubules 
a) After isolation renal stem/progenitor cells (s/pC) are placed between

layers of polyester fleece (PF) to create an artificial interstitium.

The fleece layers are mounted in a Minusheet tissue carrier.

b) For culture the carrier is transferred to a perfusion container with horizontal flow characteristics.

c) Perfusion culture is performed for 13 days under atmospheric air.

During this period always fresh culture medium is transported by a peristaltic pump

(1.25 ml/h) from a storage bottle to the container and then to awaste bottle.

案例1.干细胞体外研究，此研究应用细胞3D培养系统可以模拟体内的正常情况，干细胞疗法应用于多种疾病，

但是植入的干细胞往往被体内流体伤害，改变了细胞外基质，为了研究细胞的的适应参数，更好的起到治疗的作用，

采用了体外灌流培养系统，模拟体内体内的生理环境。此研究利用细胞灌流培养技术体外研究肾小管细胞的再生机制。

（Minuth and Denk BMC Clinical Pathology 2014, 14:34）

Reconstruction of Auto-Tissue-Engineered Lamellar Cornea by Dynamic Culture for Transplantation: 
A Rabbit Model
Abstract
To construct an auto-tissue-engineered lamellar cornea (ATELC) for transplantation,

based on acellular porcine cornealstroma and autologous corneal limbal explants,

a dynamic culture process, which composed of a submersion culture,

a perfusion culture and a dynamic air-liquid interface culture, was performed using

appropriate parameters. The results showed that the ATELC-Dynamic possessed histological

structure and DNA content that were similar to native lamellar cornea (NLC, p.0.05).

Compared to NLC, the protein contents of zonula occludens-1, desmocollin-2 and integrin b4

in ATELC-Dynamic reached 93%, 89% and 73%, respectively. The basal cells of ATELC-Dynamic

showed a better differentiation phenotype (K3 2 , P63 + , ABCG2 + ) compared with that of

ATELC in static air-lift culture (ATELC-Static, K3 + , P63 2 , ABCG2 2 ). Accordingly,

the cell-cloning efficiency of ATELC-Dynamic (9.7263.5%) was significantly higher than that

of ATELC-Static (2.1361.46%, p,0.05). The levels of trans-epithelial electrical resistance,

light transmittance and areal modulus variation in ATELC-Dynamic all reached those of NLC (p.0.05).

Rabbit lamellar keratoplasty showed that the barrier function of ATELC-Dynamic was intact,

and there were no signs of epithelial shedding or neovascularization. Furthermore,

the ATELC-Dynamic group had similar optical properties and wound healing processes compared with the NLC group. Thus,

the sequential dynamic culture process that was 
designed according to corneal physiological characteristics could successfully reconstruct

an auto-lamellar cornea with favorable morphological characteristics and satisfactory physiological function.
应用案例2.角膜移植是治疗眼部疾病的有效手段，但是角膜资源有限制约了临床上眼部疾病的治疗，

3D培养灌流系统模拟体内环境，为体外培养角膜实验提供了很好的工具，为角膜的培养及移植奠定了基础。

Advanced Cell Culture Techniques for cancer Drug Discovery
Carrie J. Lovitt,Todd B. Shelper and Vicky M.Avery*
Abstract: 
Human cancer cell lines are an integral part of drug discovery practices.
However, modeling the complexity of cancer utilizing these cell lines on

standard plastic substrata, does not accurately represent the tumor microenvironment.

Research into developing advanced tumor cell culture models in a three-dimensional (3D)

architecture that more prescisely characterizes the disease state have been undertaken

by a number of laboratories around the world. These 3D cell culture models are particularly

beneficial for investigating mechanistic processes and drug resistance in tumor cells.

In addition, a range of molecular mechanisms deconstructed by studying cancer cells in 3D

models suggest that
tumor cells cultured in two-dimensional monolayer conditions do not respond to cancer

therapeutics/compounds in a similar manner.

Recent studies have demonstrated the potential of utilizing 3D cell culture models in

drug discovery programs; however,

it is evident that further research is required for the development of more complex models

that incorporate the majority of the cellular and physical properties of a tumor.
应用案例2.细胞3D培养技术在药物研发方面发挥了很大的作用，2D细胞培养与体内细胞生长环境相差太多，

不能准确的反应实验的结果，通过模拟体内环境对细胞进行培养，检测细胞对药物的反应，使得检测结果更有意义，

为药物的研发，疾病的治疗提供了科研基础。
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参考文献