Preparation and functional evaluation of cell aggregates incorporating gelatin microspheres with different degradabilities
Preparation and functional evaluation of cell aggregates incorporating gelatin microspheres with different degradabilities
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DOI：
10.1002/term.1469
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发表时间：
2012-03
影响因子：
3.3
通讯作者：
Shuhei Tajima;Y. Tabata
中科院分区：
文献类型：
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作者：
Shuhei Tajima;Y. Tabata
本研究的目的是研究不同降解性明胶微球的细胞活力和生物学功能。用油包水乳化法制备明胶微球后，在140°C下进行不同时间的脱水热交联。体外降解实验表明，随着交联时间的延长，明胶微球降解缓慢。用明胶水凝胶微球培养MC3T3 - E1细胞，将其置于涂有聚乙烯醇的96孔微孔板的U底圆孔中，形成具有均匀分布的明胶微球的细胞聚集体。大量缓慢降解的明胶微球长时间停留在细胞聚集体中，同时观察到MC3T3‐E1细胞的高增殖。当被评估为有氧糖酵解的测量时，含有缓慢降解明胶微球的细胞聚集体中的MC3T3‐E1细胞的乳酸生成：葡萄糖消耗比含有快速降解明胶微球的聚集体中的MC3T3‐E1细胞低。含有缓慢降解明胶微球的MC3T3‐E1细胞聚集体的碱性磷酸酶活性和钙含量更高。有可能是明胶水凝胶微球的掺入具有缓慢降解性，使氧气和营养物质能够在更长的时间内渗透到细胞聚集体中，从而为细胞的存活、增殖和分化提供更好的培养条件。版权所有©2012 John Wiley & Sons， Ltd。
The objective of this study was to investigate the viability and biological functions of cells in their aggregates incorporating gelatin microspheres with different degradabilities. After being prepared by a water‐in‐oil emulsion procedure, the gelatin microspheres were dehydrothermally crosslinked at 140°C for various time periods. In vitro degradation tests showed that the gelatin microspheres were slowly degraded slowly with an increase in the crosslinking time. When MC3T3‐E1 cells were cultured with the gelatin hydrogel microspheres in the round U‐bottom wells of 96‐well microplates which had been coated with poly(vinyl alcohol), cell aggregates with homogeneously distributed gelatin microspheres were formed. A large amount of slowly degraded gelatin microspheres remained in the cell aggregates for long time periods, while a higher proliferation of MC3T3‐E1 cells was observed. When evaluated as a measure of aerobic glycolysis, the ratio of l‐lactic acid production:glucose consumption of MC3T3‐E1 cells was lower for MC3T3‐E1 cells in the cell aggregates incorporating slowly degraded gelatin microspheres than for aggregates incorporating rapidly degraded ones. The alkaline phosphatase activity and calcium content of MC3T3‐E1 cells were higher for cell aggregates incorporating slowly degraded gelatin microspheres. It is possible that the incorporation of gelatin hydrogel microspheres with slow degradability enabled the permeation of oxygen and nutrients into the cell aggregates for longer time periods, resulting in better culture conditions for the survival, proliferation and differentiation of the cells. Copyright © 2012 John Wiley & Sons, Ltd.