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基因工程和单克隆抗体技术的发展极大地促进了对细胞培养新技术的研究。为了研究出一种简单、经济的细胞培养系统，生物技术的一个新领域诞生了, 美国FiberCell系统有限公司一直致力于提供简单易用的中空纤维细胞培养系统, 让研究人员和生物技术公司 生产不可能使用传统的细胞培养的方法大规模的生产细胞及细胞的产品。 |
全新的大规模细胞培养方式--Hollow Fiber细胞培养
中空纤维细胞培养新技术-新工具--可提高100倍的产量,节省20倍的耗材.
FiberCell中空纤维细胞培养系统 利用hollow fiber membrane隔离细胞与细胞培养液，在中空纤维的外壁（exra-capillary space, ECS）培养细胞的裝置。此裝置主要為模擬生物體循環系統中毛纤维的结构及功能；由具半透析性之多孔膜狀高分子-- polysulfone天然亲水聚合物，拉成兩端有開口的纤维。將此種中空纤维裝入柱狀的塑胶容器中，其成品就像光纤排列在电缆中一样。
AHMSIC Reasearch 2010 _FiberCell中空纤维细胞培养系统_.pdf;AHMSIC (FiberCell System 12 2009).pdf; New Developments in Hollow-Fiber.pdf
应用
- 单克隆抗体试验生产或量产Monoclonal Antibodies:
每月可生产100 mg-2000 mg抗体，比用培养瓶生产抗体的浓度高100倍，可达到0.5-5 mg/mL；
- 重组蛋白表达或量产Recombinant Proteins:
每天可生产1-10 mg重组蛋白，生产分泌蛋白的表达量是培养瓶培养的100倍，可达到100-500μg/mL；
- 体外毒性研究与分析In Vitro Toxicology; 检验抗癌、抗病菌（包含HIV/HBV/HCV）以及抗寄生虫等药物反应、研究及生物测试( Bioassays)；
- 条件培养液和细胞因子的生产Conditioned Medium & Cytokine Production;
- 内皮细胞/肌细胞培养及形态研究Endothelial Cells/Muscle Cells:
在纤维内培养内皮细胞的同时，可在纤维外层培养另一种细胞（如血管平滑肌细胞）。纤维内表面积相当于一个T75培养瓶，一个培养筒可提取约100mg的RNA量。并可通过调整剪切力大小诱导内皮细胞发生形态改变，模拟体内正常与异常血压；
- 病毒扩增: 运用Virus Production可生产超过1-3×1013病毒颗粒，相当于20个滚瓶的产量；
- 淋巴细胞扩增培养及研究Lymphocyte Expansion:
细胞培养浓度可达1×108+/mL；
- 病原体培养Malaria Culture
一次培养可收获相当于60个T25培养瓶的产量。
- 人工器官研究Artificial Organ Research;
- 干细胞培养与其他细胞外基质和细胞因子培养
Stem cell and other cultures where extra-cellular matrix and cytokines can be important.
系统特点:
1. FiberCell使用波浪状的Polysulfone PlusTM中空纤维，波浪状可确保筒式培养系统内间隙一致，中空结构使其表面及周围皆可供细胞生长，纤维大量表面积还可供养分交换，是附着型与悬浮型细胞最佳培养系统！
2. 专有的正压式可置换蠕动泵系统可延长筒式培养系统的寿命，同时也能加速纤维间养分及代谢物的交换。
3. 代谢物及抑制因子可自细胞中分离出。乳酸和葡萄糖等小分子可轻易地穿过纤维。单株抗体和蛋白等大分子会被保留在毛细管间隙里。
4. 中空纤维可被活化并让重组蛋白、抗体及生长因子结合于其表面，允许长期培养以检测细胞外间质(extra-cellular matrix)的影响及生物活性的研究。
5. 亲水性的Polysulfone纤维的过滤率较传统方式（cellulosic fiber）高出10倍，可增加细胞的存活率及优化生长状态。
6. 封闭的生物安全系统可避免危险性生物对人体的威胁（P3实验室适用）。
7. 操作简便、耗材用量少、可降低血清需求量：
完整组装的无菌耗材可立即使用。
· 一天只需几分钟即可简易操作大量的细胞。
· 可进行为期六个月以上的长期细胞培养，而不需更换耗材
· 一组中空纤维细胞培养系统的产量相当于20瓶滚瓶产量
· 收取体积小，提高了抗体及蛋白质的浓度，方便纯化。
· 细胞密度高，可快速、一致的研究感染性病毒和其它微生物病原。
筒式培养系统规格：
CARTRIDGES
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SMALL Cartridge 4300-C2025 0.1µm pore size for the highest exchange rates. Activated fiber for attachment of matrix proteins, cytokines and antibodies. Ideal fiber for endothelial cell and hepatocyte culture. |
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MEDIUM Cartridges 4300-C2008 Low MWCO (5kd@50%) hydrophilic fiber for trapping smaller molecules. Suggested for recombinant proteins between 25kd and 100kd. Appropriate for suspension cell lines including CHO, HeLa and 293. Can support up to 10 9 cells and produce 100µg/mL of recombinant protein in 15mL ECS. |
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4300-C2011 High MWCO (20kd@50%) hydrophilic fiber for trapping larger molecules, hybridoma culture and lymphocyte culture. 20kd MWCO allows TGF beta and TNF alpha to diffuse away while retaining antibodies. Can support up to 10 9 cells and produce 5-50mg of monoclonal antibody every two days. Also used for medium scale adenovirus production at levels of 1- 5 x 10 12 pfu (plaque forming units).
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LARGE Cartridges 4300-C2003 Low MWCO (5kd@50%) hydrophilic fiber for trapping smaller molecules. Suggested for recombinant proteins between 25kd - 100kd. |
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4300-C2018 High MWCO (20kd@50%) hydrophilic fiber for trapping larger molecules, hybridoma and lymphocyte cultures. 20kd MWCO allows TGF beta and TNF alpha to diffuse away while retaining antibodies. Can support up to 10 11 cells and produce 75-150mg of monoclonal antibody every two days. Appropriate for adherent suspension cell lines including CHO, HeLa and 293 cells. Can support up to 10 11 cells and produce 100µg/mL of recombinant protein in 70mL ECS.
X-LARGE Cartridges
4300-C4005 The C4005 cartridge is intended for use in larger hollow fiber cell culture systems from other manufacturers and does not include a flow path stand or oxygenator tubing. Side ports have 3" of tubing capped with luer fittings, end ports are 3/8" hose barbs. High gross filtration rate and polysulfone fiber are superior to cellulose acetate for recombinant protein production.
4300-C4020 The C4020 cartridge is intended for use in larger hollow fiber cell culture systems from other manufacturers and does not include a flow path stand or oxygenator tubing. Side ports have 3" of tubing capped with luer fittings, end ports are 3/8" hose barbs with tubing and standard luer connectors on the ends. High gross filtration rate and polysulfone fiber are superior to cellulose acetate for recombinant protein and monoclonal antibody production.
Cartridge/Flowpath Specifications
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Stock No. |
Size |
Surface Area |
Fiber Type |
Packing Density |
ECS Vol |
MWCO 50% |
Max. Cell# |
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C2025 |
Small |
75cm2 |
activated PS |
30% |
2.5mL |
0.1µm |
108 |
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C2008 |
Medium |
3000cm2 |
low flux PS |
50% |
15mL |
5kd |
109 |
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C2011 |
Medium |
3000cm2 |
high flux PS |
50% |
15mL |
20kd |
109 |
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C2003 |
Large |
1.2m2 |
low flux PS |
50% |
70mL |
5kd |
5 x 1010 |
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C2018 |
Large |
1.2m2 |
high flux PS |
50% |
70mL |
20kd |
5 x 1010 |
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C4005 |
X-Large |
2.5m2 |
low flux PS |
50% |
150mL |
5kd |
1011 |
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C3008 |
Medium |
2000cm2 |
Cellulosic |
38% |
12mL |
10kd |
1011 |
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C4020 |
X-Large |
2.5m2 |
high flux PS |
50% |
150mL |
20kd |
1011 |
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C5011 |
Medium |
2100cm2 |
high flux PS |
50% |
15mL |
20kd |
2 x 1010 |
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Oxy0.6 |
80mls |
6000cm2 |
Hydrophobic Polyethylene |
N/A |
40mL |
100kd |
N/A |
FiberCell Systems Cartridges (except X-Large) are supplied assembled with the flow path ready to use. They are pre-sterilized and double bagged with instructions provided.
Advantages of Hollow Fiber Cell Culture
- Provides 10X the gross filtration rate of cellulosic hollow fibers.
- Improves cell viability.
- Eliminates dead spots with uniform fiber spacing and "wavy" fibers within the cartridge.
- Dialyzes waste and inhibitory cytokines away from cells.
- Stabalizes the PH of the cell culture medium.
- Ehances bio-safety with closed system.
- Handles large cell numbers (up to 10 11)
- Concentrates secreted proteins and antibodies in the small volume of the extra-capillary space.
- Provides flexible control options with unique micro-processor controlled pump assembly.
- Ensures long life of the cartridge and facilitates nutrient and waste exchange across the fiber with positive-pressure displacement pumping system.