Effects of heat treatment and concentration of fish serum on cell growth in adhesion culture of Chinese hamster ovary cells

Fujiwara, Masashi; Tsukada, Ryohei; Shioya, Itaru; Takagi, Mutsumi

doi:10.1007/s10616-009-9199-7

Cited by 5 publications

(4 citation statements)

References 9 publications

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“…Several groups already demonstrated that differences in serum concentration or type could highly affect cell adhesion (Fujiwara et al, 2009;Grinnell and Feld, 1979;Schmidt et al, 2011). Our study confi rmed that serum greatly affects cell adhesion in addition to surface patterning.…”

Section: Wwwecmjournalorg E Lamers Et Al Dynamic Cell Adhesion Andsupporting

confidence: 81%

Dynamic cell adhesion and migration on nanoscale grooved substrates

Lamers

Riet²,

Domański³

et al. 2012

eCM

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Organised nanotopography mimicking the natural extracellular matrix can be used to control morphology, cell motility, and differentiation. However, it is still unknown how specifi c cell types react with specifi c patterns. Both initial adhesion and preferential cell migration may be important to initiate and increase cell locomotion and coverage with cells, and thus achieve an enhanced wound healing response around an implantable material. Therefore, the aim of this study was to evaluate how MC3T3-E1 osteoblast initial adhesion and directional migration are infl uenced by nanogrooves with pitches ranging from 150 nm up to 1000 nm. In this study, we used a multipatterned substrate with fi ve different groove patterns and a smooth area with either a concentric or radial orientation. Initial cell adhesion measurements after 10 s were performed using atomic force spectroscopy-assisted single-cell force spectroscopy, and demonstrated that nascent cell adhesion was highly induced by a 600 nm pitch and reduced by a 150 nm pitch. Addition of RGD peptide signifi cantly reduced adhesion, indicating that integrins and cell adhesive proteins (e.g. fi bronectin or vitronectin) are key factors in specifi c cell adhesion on nanogrooved substrates. Also, cell migration was highly dependent on the groove pitch; the highest directional migration parallel to the grooves was observed on a 600 nm pitch, whereas a 150 nm pitch restrained directional cell migration. From this study, we conclude that grooves with a pitch of 600 nm may be favourable to enhance fast wound closure, thereby promoting tissue regeneration.

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Section: Wwwecmjournalorg E Lamers Et Al Dynamic Cell Adhesion Andsupporting

confidence: 81%

Dynamic cell adhesion and migration on nanoscale grooved substrates

Lamers

Riet²,

Domański³

et al. 2012

eCM

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“…The cell-growth-stimulating activity of fish serum for MSCs is yet unclear, although the use of fish serum has been investigated (Fujiwara et al 2007(Fujiwara et al , 2009(Fujiwara et al , 2010. The in vitro cell-growth-stimulating activity of adult human serum was much lower than that of fetal serum.…”

mentioning

confidence: 99%

Xeno-free proliferation of human bone marrow mesenchymal stem cells

et al. 2011

Self Cite

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The proliferation of human bone marrow mesenchymal stem cells (MSCs) employing xeno-free materials not containing fetal calf serum (FCS) and porcine trypsin was investigated for the regenerative medicine of cartilage using MSCs. Four sequential subcultivations of MSCs using a medium containing 10% FCS and recombinant trypsin (TrypLESelect™) resulted in cell growth comparable to that with porcine trypsin. There was no apparent difference in the cell growth and morphology between two kinds of MSC stored in liquid nitrogen using 10% FCS plus DMSO or serum-free TC protector™. MSCs were isolated from human bone marrow cells, stored in liquid nitrogen, and sequentially subcultivated four times employing conventional materials that included FCS, porcine trypsin, and DMSO, or xeno-free materials that included serum-free medium (MesenCult-XF™), TC protector™ and TrypLESelect™. Cells in the culture using the xeno-free materials maintained typical fibroblast-like morphology and grew more rapidly than the cells in the culture using the conventional materials, while the cell surface markers of MSCs (CD90 and CD166) were well maintained in both cultures. Chondrogenic pellet cultures were carried out using these subcultivated cells and a medium containing TGFβ3 and IGF1. The pellet culture using cells grown with the xeno-free materials showed an apparently higher gene expression of aggrecan, a chondrocyte marker, than the pellet culture using cells grown with the conventional materials. Consequently, MSCs that are isolated, stored, and grown using the xeno-free materials including the serum-free medium (MesenCult-XF™), TC protector™, and recombinant trypsin (TrypLESelect™) might be applicable for regenerative medicine of cartilage.

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“…The released cells were mix of plenty of myoblasts and fibroblasts, and the external characteristics of both types of cells were so similar that they cannot be distinguished and well separated by their external characteristics. However, the fibroblasts could adhere on some plastic surface more easily than the myoblasts did (Fujiwara et al 2009). Thus, we obtained plenty of duck myoblasts with high purity by using the differential attachment technique.…”

Section: Discussionmentioning

confidence: 99%

“…The proliferation and differentiation of myoblasts can be influenced by the choice of the serum concentration in the culture medium. Low concentration of serum in culture medium can induce cell differentiation, whereas high concentration serum can lead to a proliferation tendency of the cells (Fujiwara et al 2009). In this study, we added serum with a concentration of 15% to the culture medium, resulting in a longer proliferation phase for the duck myoblasts.…”

Section: Discussionmentioning

confidence: 99%

Characterization of in vitro cultured myoblasts isolated from duck (Anas platyrhynchos) embryo

Liu

Chen

et al. 2011

Cytotechnology

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Myoblasts isolated from duck embryonic muscle were purified and in vitro cultured. External characteristics were observed by using the immunofluorescence technique, and growth curve of duck embryonic myoblasts was established after measuring with the MTT method. Moreover, mRNA expression of three marker genes, the Desmin, the muscle creatine kinase (Mck) and the troponin C (Tnnc), which could reflect the development status of myofibers, were detected each 24 h for cultured cells by using the qPCR technique. Results showed that the in vitro cultured duck myoblasts went through a series of developmental stages, including the proliferation of myoblasts, the differentiation of multi-nuclei myotubes, and the formation of myofiber. The cultured duck embryonic myoblasts entered into a logarithmic stage approximately on the fourth day after seeding. Accompanying with its progressive growth before entering into the logarithmic phase, the myoblasts also showed some differentiation phenomena, reflected by a low expression level of Desmin and high expression level of the Mck and Tnnc genes. During the rapid growth of the logarithmic phase, there was a high expression of the Desmin gene, and a low expression level of the Mck gene and the Tnnc gene in the cultured myoblasts. The expression profiles of the three marker genes for muscle development could be used for distinguishing the different developmental stages of in vitro cultured myoblasts at the molecular level, which would be more accurate and more feasible than observing the external characteristics of the cultured cells.

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Effects of heat treatment and concentration of fish serum on cell growth in adhesion culture of Chinese hamster ovary cells

Cited by 5 publications

References 9 publications

Dynamic cell adhesion and migration on nanoscale grooved substrates

Dynamic cell adhesion and migration on nanoscale grooved substrates

Xeno-free proliferation of human bone marrow mesenchymal stem cells

Characterization of in vitro cultured myoblasts isolated from duck (Anas platyrhynchos) embryo

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