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- W4387091965 abstract "Feeder cells play a crucial role in maintaining the pluripotency of embryonic stem cells (ESCs) by secreting various extrinsic regulators, such as extracellular matrix (ECM) proteins and growth factors. Although primary mouse embryonic fibroblasts (MEFs) are the most widely used feeder cell type for the culture of ESCs, they have inevitable disadvantages such as batch-to-batch variation and labor-intensive isolation processes. Here, we revealed that the Sandoz inbred Swiss mouse (SIM) thioguanine-resistant ouabain-resistant (STO) cell line, an immortalized cell line established from mouse SIM embryonic fibroblasts, can be used as a feeder layer for in vitro culture of authentic pig ESCs instead of primary MEFs. First, the expression of genes encoding ECM proteins and growth factors was analyzed to compare their secretory functions as feeder cells. Quantitative real-time PCR showed that the gene expression of these pluripotency-associated factors was downregulated in STO cells compared to primary MEFs of similar density. Therefore, subsequent optimization of the culture conditions was attempted using higher STO cell densities. Notably, pig ESCs cultured on STO cell density of 3X (187,500 cells/cm2) exhibited the most similar pluripotent state to pig ESCs cultured on primary MEF density of 1X (62,500 cells/cm2), as determined by alkaline phosphatase staining, quantitative real-time PCR, and immunocytochemistry. Additionally, pig ESCs cultured on STO cell density of 3X formed complex teratoma containing multiple types of tissues derived from all three germ layers. Our culture conditions using optimal STO cell density can be applied to fields requiring reproducible and scalable production of pig ESCs, such as preclinical research and cellular agriculture." @default.
- W4387091965 created "2023-09-28" @default.
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- W4387091965 date "2023-09-27" @default.
- W4387091965 modified "2023-10-05" @default.
- W4387091965 title "Development of reproducible and scalable culture conditions for in vitro maintenance of pig Embryonic stem cells using the Sandoz inbred Swiss mouse thioguanine-resistant ouabain-resistant cell line as a feeder layer" @default.
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- W4387091965 doi "https://doi.org/10.1089/scd.2023.0171" @default.
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