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TGF-3 pretreatment can also improve the survival and matrix accumulation of BMSCs in acidic environments ( Notably, the duration and degree of acidic exposure may have dual effects: short-term acidic treatment (pH 6.56.8, 17 days) can enhance the stemness of BMSCs, promoting spheroid colony formation (Wuertz et al., 2008

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Even if you follow a protocol to the letter, understanding why you're doing something builds intuition in the lab