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This was achieved by serially passaging viable cells for multiple passages.
hES cells (approximately 2×106 cells) grown under animal-derived extracellular matrices-free culture conditions for multiple passages with occasional freezing and thawing were subcutaneously injected into severe combined immunodeficient (SCID / beige mice (Charles River Laboratories, Wilmington, MA).
Similar to "good" preparations of manually isolated NSC, these cultures were often free of NSC contaminants and could be cultured in vitro for multiple passages (up to 22 tested).
Generating homozygotes in F1 transgenics using a "mutagenic chain reaction" (Gantz and Bier 2015), would limit the need for multiple passages and mating to establish a line.
By routinely propagating lines at high density and passaging with trypsin, we can maintain cell lines with normal karyotype for multiple passages.
These cells were able to grow as a monolayer on Matrigel-coated dishes and could be grown for multiple passages in the presence of bFGF and EGF.
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For instance, multiple passages of the cell lines IMR90 (human fetal lung fibroblast) and H1 (human embryonic stem cells) showed high concordance [ 17].
The current data argue to the contrary, suggesting that the amplification and deletions seen in cell lines offer the same growth advantages in cell culture that they do in vivo, and are similarly selected for over multiple passages.
Therefore, Enterobacter agglomerans was isolated from mosquitoes and selected for midgut survival by multiple passages through mosquitoes.
The re-differentiated fibroblast-like lines (TER lines) were produced from explants of IPSCA and B teratomas, expanded in vitro, and selected over multiple passages for cells of fibroblast morphology.
Another disadvantage for BCCs subjected to multiple passages relates to their genomic instability resulting in a population of cells with varying degrees of tumorigenic potential.
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