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Food-Derived Collagen Peptides, Prolyl-Hydroxyproline (Pro-Hyp), and Hydroxyprolyl-Glycine (Hyp-Gly) Enhance Growth of Primary Cultured Mouse Skin Fibroblast Using Fetal Bovine Serum Free from Hydroxyprolyl Peptide

1
Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto 606 8502, Japan
2
Department of Food Science and Nutrition, Faculty of Human Life and Environment, Nara Women’s University, Nara 630 8506, Japan
3
Division of Applied Life Sciences, Graduate School of Life and Environment Sciences, Kyoto Prefectural University, Kyoto 606 8522, Japan
4
Peptide Division, Nitta Gelatin Inc., 2-22 Futamata, Yao, Osaka 581 0024, Japan
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(1), 229; https://doi.org/10.3390/ijms21010229
Received: 30 November 2019 / Revised: 25 December 2019 / Accepted: 26 December 2019 / Published: 28 December 2019
(This article belongs to the Special Issue Peptides for Health Benefits 2019)
Prolyl-hydroxyproline (Pro-Hyp) and hydroxyprolyl-glycine (Hyp-Gly) appear in human blood after ingestion of collagen hydrolysate and trigger growth of fibroblasts attached on collagen gel, which has been associated with beneficial effects upon ingestion of collagen hydrolysate, such as improvement of skin and joint conditions. In the present study, inconsistent results were obtained by using different lots of fetal bovine serum (FBS). Fibroblasts proliferated in collagen gel without adding Pro-Hyp and Hyp-Gly and did not respond to addition of Pro-Hyp and Hyp-Gly, which raises doubts about conclusions from prior research. Unexpectedly high levels of hydroxyprolyl peptides, including Pro-Hyp, however, were present in the FBS (approximately 100 µM), and also in other commercially available forms of FBS (70–80 µM). After removal of low molecular weight (LMW, < 6000 Da) compounds from the FBS by size exclusion chromatography, Pro-Hyp and Hyp-Gly again triggered growth of fibroblasts attached on collagen and increased the number of fibroblasts migrated from mouse skin. These results indicate the presence of bioactive hydroxyprolyl peptides in commercially available FBS, which can mask effects of Pro-Hyp and Hyp-Gly supplementation; our work confirms that Pro-Hyp and Hyp-Gly do play crucial roles in proliferation of fibroblasts. View Full-Text
Keywords: prolyl-hydroxyproline (Pro-Hyp); hydroxyprolyl-glycine (Hyp-Gly); collagen peptide; fibroblasts; fetal bovine serum (FBS) prolyl-hydroxyproline (Pro-Hyp); hydroxyprolyl-glycine (Hyp-Gly); collagen peptide; fibroblasts; fetal bovine serum (FBS)
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Asai, T.T.; Oikawa, F.; Yoshikawa, K.; Inoue, N.; Sato, K. Food-Derived Collagen Peptides, Prolyl-Hydroxyproline (Pro-Hyp), and Hydroxyprolyl-Glycine (Hyp-Gly) Enhance Growth of Primary Cultured Mouse Skin Fibroblast Using Fetal Bovine Serum Free from Hydroxyprolyl Peptide. Int. J. Mol. Sci. 2020, 21, 229.

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