Fibroblast phenylalanine concentration as a surrogate biomarker of cellular number
Por:
Oliva-Mussara C, Arias A, Ruiz M, Pujol A, Garrabou G, Canto-Santos J, Urreizti R, Castilla-Vallmanya L, Rodriguez H, Jou-Munoz C, Casado-Rio M, Ormazabal-Herrero A and Artuch-Iriberri R
Publicada:
15 jul 2023
Ahead of Print:
1 jun 2023
Resumen:
Metabolomics studies in human dermal fibroblasts can elucidate the biological mechanisms associated with some diseases, but several methodological issues that increase variability have been identified. We aimed to quantify the amino acid levels in cultured fibroblasts and to apply different sample-based normalization approaches. Forty-four skin biopsies from control subjects were collected. Amino acids were measured in fibroblasts supernatants by UPLC-MS/MS. Statistical supervised and unsupervised studies were used. Spearman's test showed that phenylalanine displayed the second highest correlation with the remaining amino acids (mean r = 0.8), whereas the total protein concentration from the cell pellet showed a mean of r = 0.67. The lowest percentage of variation was obtained when amino acids were normalized by phenylalanine values, with a mean of 42% vs 57% when normalized by total protein values. When amino acid levels were normalized by phenylalanine, Principal Component Analysis and clustering analyses identified different fibroblasts groups. In conclusion, phenylalanine may be a suitable biomarker to estimate cellular content in cultured fibroblasts.
Filiaciones:
Oliva-Mussara C:
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Arias A:
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Ruiz M:
Neurometabolic Diseases Laboratory, Bellvitge Biomedical Research Institute (IDIBELL), Avinguda de la Gran via de l'Hospitalet, 199, 08908 L'Hospitalet de Llobregat, Barcelona, Spain
Catalan Institution of Research and Advanced Studies (ICREA), Universitat Autònoma de Barcelona, Av. de Serragalliners, s/n, 08193 Bellaterra, Barcelona, Spain
Pujol A:
Neurometabolic Diseases Laboratory, Bellvitge Biomedical Research Institute (IDIBELL), Avinguda de la Gran via de l'Hospitalet, 199, 08908 L'Hospitalet de Llobregat, Barcelona, Spain
Catalan Institution of Research and Advanced Studies (ICREA), Universitat Autònoma de Barcelona, Av. de Serragalliners, s/n, 08193 Bellaterra, Barcelona, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Garrabou G:
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Muscle Research and Mitochondrial Function Laboratory, CELLEX-August Pi i Sunyer Biomedical Research Institute, C/ del Roselló, 149, 08036, Barcelona, Spain
Faculty of Medicine and Health Sciences-University of Barcelona, C/ de Casanova, 143, 08036 Barcelona, Spain
Internal Medicine Department-Hospital Clinic of Barcelona, C/ del Roselló, 149, 08036 Barcelona, Spain
Canto-Santos J:
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Muscle Research and Mitochondrial Function Laboratory, CELLEX-August Pi i Sunyer Biomedical Research Institute, C/ del Roselló, 149, 08036, Barcelona, Spain
Urreizti R:
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Castilla-Vallmanya L:
Department of Genetics, Microbiology and Statistics, Institute of Biomedicine (IBUB), Faculty of Biology, Universitat de Barcelona, Avinguda Diagonal, 643, 08028 Barcelona, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Rodriguez H:
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Jou-Munoz C:
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Biobank and Pathology department, Hospital Sant Joan de Deu, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Casado-Rio M:
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Ormazabal-Herrero A:
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Artuch-Iriberri R:
Clinical Biochemistry Department, Sant Joan de Déu Research Institute, Passeig Sant Joan de Déu, 2, 08950, Esplugues de Llobregat, Barcelona, Spain
Centro de Investigación Biomédica en Red de Enfermedades Raras, ISCIII, Avda. de Monforte de Lemos, 5, 28029 Madrid, Spain
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