Next generation in vitro liver model design: Combining a permeable polystyrene membrane with a transdifferentiated cell line.
Next generation in vitro liver model design: Combining a permeable polystyrene membrane with a transdifferentiated cell line.
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DOI：
10.1016/j.memsci.2018.07.063
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发表时间：
2018-11-01
影响因子：
9.5
通讯作者：
Ellis MJ
中科院分区：
文献类型：
--
作者：
Luetchford KA;Wung N;Argyle IS;Storm MP;Weston SD;Tosh D;Ellis MJ
在这里，我们描述了生物相容性，多孔聚苯乙烯膜，适用于细胞培养的制造和表征。聚苯乙烯虽然在传统的细胞培养中得到了广泛的应用，但由于其疏水性和无孔结构，尚未被用作中空纤维膜。在这里，我们使用微晶氯化钠（4.7 ± 1.3 µm）来控制聚苯乙烯膜的孔隙度，并使用氧等离子体表面处理来降低疏水性。孔隙素浓度的增加与表面孔隙密度、大孔隙形成、渗透率和平均孔径的增加有关，但与机械强度的降低有关。对于组织工程应用，从含有40% （w/w）氯化钠的铸造溶液中纺出的膜代表了强度和渗透性之间的平衡，表面孔隙密度为208.2 ± 29.7孔/mm2，平均表面孔径为2.3 ± 0.7 µm，杨氏模量为115.0 ± 8.2 MPa。我们通过令人兴奋的细胞系-培养基组合证明了该材料的生物相容性：AR42J-B13胰腺细胞系转分化为肝细胞样细胞。用地塞米松/癌他汀- m治疗AR42J-B13超过14天可诱导向肝脏表型的转分化。胰腺表型明显丧失，表现为胰腺标记淀粉酶的表达丧失，而肝脏表型增加，表现为肝脏标记转铁蛋白、氨基甲酰磷酸合成酶和谷氨酰胺合成酶的表达诱导。这种膜制造方法与转分化肝细胞生物相容性的证明相结合，为体外肝模型和生物人工肝装置提供了一种新颖、优越的替代设计。膜的生产条件温和，经济可行。微晶氯化钠可以控制不对称膜的孔隙率。生物相容性非可生物降解膜支持细胞活力和转分化。转分化的AR42J-B13细胞有可能用于体外肝脏模型。
Herein we describe the manufacture and characterisation of biocompatible, porous polystyrene membranes, suitable for cell culture. Though widely used in traditional cell culture, polystyrene has not been used as a hollow fibre membrane due to its hydrophobicity and non-porous structure. Here, we use microcrystalline sodium chloride (4.7 ± 1.3 µm) to control the porosity of polystyrene membranes and oxygen plasma surface treatment to reduce hydrophobicity. Increased porogen concentration correlates to increased surface pore density, macrovoid formation, gas permeability and mean pore size, but a decrease in mechanical strength. For tissue engineering applications, membranes spun from casting solutions containing 40% (w/w) sodium chloride represent a compromise between strength and permeability, having surface pore density of 208.2 ± 29.7 pores/mm2, mean surface pore size of 2.3 ± 0.7 µm, and Young's modulus of 115.0 ± 8.2 MPa. We demonstrate the biocompatibility of the material with an exciting cell line-media combination: transdifferentiation of the AR42J-B13 pancreatic cell line to hepatocyte-like cells. Treatment of AR42J-B13 with dexamethasone/oncostatin-M over 14 days induces transdifferentiation towards a hepatic phenotype. There was a distinct loss of the pancreatic phenotype, shown through loss of expression of the pancreatic marker amylase, and gain of the hepatic phenotype, shown through induction of expression of the hepatic markers transferrin, carbamoylphosphate synthetase and glutamine synthetase. The combination of this membrane fabrication method and demonstration of biocompatibility of the transdifferentiated hepatocytes provides a novel, superior, alternative design for in vitro liver models and bioartificial liver devices. The conditions of membrane production are mild and economically viable. Microcrystalline sodium chloride enables controlled porosity of asymmetric membranes. Biocompatible non-biodegradable membranes support cell viability and transdifferentiation. Transdifferentiated AR42J-B13 cells offer potential for use in in vitro liver models.
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影响因子：
21.3
作者：
Shen, CN;Slack, JMW;Tosh, D
通讯作者：
Tosh, D
影响因子：
4
作者：
Ajioka, I;Akaike, T;Watanabe, Y
通讯作者：
Watanabe, Y
影响因子：
5.6
作者：
Burke, ZD;Shen, CN;Tosh, D
通讯作者：
Tosh, D
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3
作者：
FROMMER, MA;SHPORER, M;MESSALEM, RM
通讯作者：
MESSALEM, RM
影响因子：
3.7
作者：
Li, K;Kong, JF;Teo, WK
通讯作者：
Teo, WK