共查询到20条相似文献,搜索用时 31 毫秒
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Li WC Ralphs KL Slack JM Tosh D 《The international journal of biochemistry & cell biology》2007,39(3):541-554
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Xiao-xi Xu Chang Liu Yang Liu Nan Li Xin Guo Shu-jun Wang Guang-wei Sun Wei Wang Xiao-jun Ma 《Experimental cell research》2013
Hepatocellular carcinoma (HCC) is the most common primary liver cancer and often forms metastases, which are the most important prognostic factors. For further elucidation of the mechanism underlying the progression and metastasis of HCC, a culture system mimicking the in vivo tumor microenvironment is needed. In this study, we investigated the metastatic ability of HCC cells cultured within alginate gel (ALG) beads. In the culture system, HCC cells formed spheroids by proliferation and maintained in nuclear abnormalities. The gene and protein expression of metastasis-related molecules was increased in ALG beads, compared with the traditional adhesion culture. Furthermore, several gene expression levels in ALG bead culture system were even closer to liver cancer tissues. More importantly, in vitro invasion assay showed that the invasion cells derived from ALG beads was 7.8-fold higher than adhesion cells. Our results indicated that the in vitro three-dimensional (3D) model based on ALG beads increased metastatic ability compared with adhesion culture, even partly mimicked the in vivo tumor tissues. Moreover, due to the controllable preparation conditions, steady characteristics and production at large-scale, the 3D ALG bead model would become an important tool used in the high-throughput screening of anti-metastasis drugs and the metastatic mechanism research. 相似文献
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Production of recombinant human acetylcholinesterase (AChE) by a high producer human embryonic kidney cell line (293) was evaluated by three main cell propagation systems; surface propagator, fixed-bed reactor and stirred microcarrier cultures. The recombinant cell line expresses AChE levels as high as 10–20 mg/l/day. System productivities in either the surface propagator (multitray system), or in the fixed-bed reactor (polyurethane macroporous sponges) were 4–8 mg AChE/l/day during a production period of 8 days. Similar productive rates, yet longer production periods (up to 22 days), were obtained in microcarrier (MC) cultures using either polystyrene beads (Biosilon); collagen-coated dextran beads (Cytodex-3); or gelatin macroporous beads (Cultispher-G). Best results were obtained in an aggregate cculture using cellulose beads charged with diethylaminoethyl (DEAE) groups, (Servacel), as carriers. In this culture, a system productivity of 6–10 mg/l/day was maintained for 28 days. 相似文献
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Calcium-alginate gel bead cross-linked with gelatin as microcarrier for anchorage-dependent cell culture 总被引:4,自引:0,他引:4
Valuable products obtainedfrom the cultivation of anchorage-dependent mammalian cells require large-scale processes to obtain commercially useful quantities. It is generally accepted that suspension culture is the ideal mode of operation. Because anchorage-dependent cells need surfaces to be able to attach and spread, the incorporation of microcarriers to suspension culture is indispensable. Since the dextran-based microcarrier wasfirst introduced, many different types of microcarriers have been developed and commercialized. In this study, alginate-based microcarriers were made in the following order: (i) calcium-alginate gel beads prepared by dropping a blend of sodium alginate and propylene glycol alginate (PGA) into calcium chloride solution, (ii) the PGA section of gel beads cross-linked with gelatin in alkaline solution (i.e., via the transacylation reaction between the ester group of PGA and amino group of gelatin), and (iii) gelatin membrane around the beads further cross-linked by glutaraldehyde. The glutaraldehyde-treated gelatintransacylated PGA/alginate microcarrier showed superior features in high stability under phosphate-containing solution, density close to that of culture medium, and transparency. Moreover, the Chinese hamster ovary CHO-KI and amphotropic retrovirus producer PA317 cells cultivated on the newly synthesized microcarriers exhibited similar growth kinetics of these two types of cell lines cultured on commercial polystyrene microcarriers. However, cell morphology was easily monitored on the transparent microcarriers made in this study. 相似文献
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Pure gelatin microcarriers: Synthesis and use in cell attachment and growth of fibroblast and endothelial cells 总被引:1,自引:0,他引:1
Kimberly W. Wissemann Bruce S. Jacobson 《In vitro cellular & developmental biology. Plant》1985,21(7):391-401
Summary A new type of microcarrier was described using bead emulsion-polymerization techniques. An aqueous solution of gelatin and
glutaraldehyde was dispersed in a hydrophobic phase of mineral oil, using Triton X-114 as an emulsifier, and polymerization
was initiated. The resultant spherical beads, composed entirely of gelatin, showed excellent mechanical stability to ethanol
drying, sterilization, and long-term use in microcarrier spinner cultures. The solid gelatin microcarriers supported the growth
of L-929 fibroblast, swine aorta endothelial, human umbilical endothelial, and HeLa-S3 cultures with no adverse effects on cell morphology or growth. The beads were transparent in growth medium and attached cells
were clearly visualized without staining. The beads were also compatible with techniques for scanning electron microscopy.
Collagenase could be used to entirely digest the gelatin beads, leaving the cells free from microcarriers and suspended in
solution while retaining 98% cell viability. The results further showed that after collagenase treatment the cells would populate
fresh gelatin microcarriers and grow to confluence. Cell attachment kinetics revealed that the endothelial cells attached
to the gelatin beads at the same rate as to tissue culture plates, whereas the fibroblast cells attached to the beads more
slowly. However, once the fibroblast cells were attached to the gelatin microcarriers they spread and grew normally.
This research was supported in part by the National Institutes of Health (GN 29127) and Ventrex Laboratories, Portland, Maine. 相似文献
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