TGF-β2, EGF and FGF21 groups were supplemented with recombinant human TGF-β2 (97.3% homology with rat), recombinant rat EGF and recombinant human FGF21 (92% homology with rat) (all from Peprotech, Rocky Hill, NJ, USA). These products were reconstituted in phosphate-buffered solution (PBS, pH 7.2) with 0.1% bovine serum albumin (BSA, SeraCare Life Sciences, Milford, MA, USA), according to the manufacturer’s recommendations. The dose of TGF-β2 was 35 μg/kg/day, which was based on the amount of TGF-β2 found in mid-lactation rat milk and milk intake by pups at 4–14 days of age [13 (link)]. The dose of EGF was 100 μg/kg/day, which was demonstrated to be effective as a treatment in a rat model of NEC [29 (link)]. Finally, the dose of FGF21 was 5 μg/kg/day, an amount that was established in relation to TGF-β2, which was found in a 1:10 ratio FGF21:TGF-β2 [23 (link),30 (link)]. The REF group received a matched volume of the vehicle (1% BSA in PBS) used to dilute the GF.
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