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不同微载体上胎儿软骨细胞的分离、鉴定和扩增。

Derivation, characterization and expansion of fetal chondrocytes on different microcarriers.

机构信息

TUBITAK MRC-Genetic Engineering Biotechnology Institute (GEBI), 41470, Gebze/Kocaeli, Turkey.

出版信息

Cytotechnology. 2011 Dec;63(6):633-43. doi: 10.1007/s10616-011-9380-7. Epub 2011 Aug 12.

Abstract

Fetal chondrocytes (FCs) have recently been identified as an alternative cell source for cartilage tissue engineering applications because of their partially chondrogenically differentiated phenotype and developmental plasticity. In this study, chondrocytes derived from fetal bovine cartilage were characterized and then cultured on commercially available Cytodex-1 and Biosilon microcarriers and thermosensitive poly(hydroxyethylmethacrylate)-poly(N-isopropylacrylamide) (PHEMA-PNIPAAm) beads produced by us. Growth kinetics of FCs were estimated by means of specific growth rate and metabolic activity assay. Cell detachment from thermosensitive microcarriers was induced by cold treatment at 4 °C for 20 min or enzymatic treatment was applied for the detachment of cells from Cytodex-1 and Biosilon. Although attachment efficiency and proliferation of FCs on PHEMA-PNIPAAm beads were lower than that of commercial Cytodex-1 and Biosilon microcarriers, these beads also supported growth of FCs. Detached cells from thermosensitive beads by cold induction exhibited a normal proliferative activity. Our results indicated that Cytodex-1 microcarrier was the most suitable material for the production of FCs in high capacity, however, 'thermosensitive microcarrier model' could be considered as an attractive solution to the process scale up for cartilage tissue engineering by improving surface characteristics of PHEMA-PNIPAAm beads.

摘要

胎儿软骨细胞 (FCs) 因其部分软骨分化表型和发育可塑性,最近被认为是软骨组织工程应用的替代细胞来源。在这项研究中,从胎牛软骨中分离出来的软骨细胞被进行了特征描述,然后在我们制备的市售 Cytodex-1 和 Biosilon 微载体以及热响应性聚(羟甲基丙烯酸乙酯)-聚(N-异丙基丙烯酰胺) (PHEMA-PNIPAAm) 珠上进行培养。通过比生长速率和代谢活性测定来估计 FCs 的生长动力学。通过在 4°C 下冷处理 20 分钟或用酶处理从 Cytodex-1 和 Biosilon 上分离细胞来诱导热敏微载体上 FCs 的细胞脱离。尽管 PHEMA-PNIPAAm 珠上 FCs 的附着效率和增殖低于商业 Cytodex-1 和 Biosilon 微载体,但这些珠也支持 FCs 的生长。通过冷诱导从热敏珠上分离的细胞表现出正常的增殖活性。我们的结果表明,Cytodex-1 微载体是生产高容量 FCs 的最适合材料,然而,“热敏微载体模型”可以通过改善 PHEMA-PNIPAAm 珠的表面特性来考虑作为软骨组织工程过程放大的有吸引力的解决方案。

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Plasticity of fetal cartilaginous cells.胎儿软骨细胞的可塑性。
Cell Transplant. 2010;19(10):1349-57. doi: 10.3727/096368910X506854. Epub 2010 May 4.
2
Expansion and Harvesting of hMSC-TERT.人端粒酶逆转录酶永生化间充质干细胞的扩增与收获
Open Biomed Eng J. 2007 Sep 7;1:38-46. doi: 10.2174/1874120700701010038.
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Bone tissue engineering using foetal cell therapy.
Swiss Med Wkly. 2006 Sep 2;136(35-36):557-60. doi: 10.4414/smw.2006.11411.
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Microcarriers in the engineering of cartilage and bone.软骨和骨组织工程中的微载体
Trends Biotechnol. 2006 Jul;24(7):299-304. doi: 10.1016/j.tibtech.2006.04.009. Epub 2006 May 4.

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