Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts
BACKGROUND Human dermal fibroblast (HDF) cultures can be used as a regenerative agent for wound healing. Fetal bovine serum (FBS) as a culture supplement is derived from animals, therefore not being constant in composition, causes variations in research results, thus requiring a substitute such as...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Faculty of Medicine Trisakti University
2023-09-01
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| Series: | Universa Medicina |
| Subjects: | |
| Online Access: | https://univmed.org/ejurnal/index.php/medicina/article/view/1476 |
| _version_ | 1797371779001352192 |
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| author | Nur Ilham Risma Hidayati Indra Kusuma Eko Poerwanto |
| author_facet | Nur Ilham Risma Hidayati Indra Kusuma Eko Poerwanto |
| author_sort | Nur Ilham Risma Hidayati |
| collection | DOAJ |
| description | BACKGROUND
Human dermal fibroblast (HDF) cultures can be used as a regenerative agent for wound healing. Fetal bovine serum (FBS) as a culture supplement is derived from animals, therefore not being constant in composition, causes variations in research results, thus requiring a substitute such as KnockOut serum replacement (KOSR). This study evaluated the defined KOSR as FBS substitute for HDF culture by measuring the relative expression of basic fibroblast growth factor (bFGF) and keratinocyte growth factor (KGF) messenger RNA (mRNA), HDF cell proliferation, and HDF migration.
METHODS
Human dermal fibroblast culture was divided into 2 intervention groups receiving KOSR 5% and KOSR 10%, respectively, and a control group receiving FBS 10%. Reverse transcription polymerase chain reaction (RT-PCR) was used for bFGF and KGF mRNA relative expression at the fifth passage (P5). Cell counting kit-8 (CCK-8) reagent was used for the HDF cell proliferation assay at P5 and the scratch assay was used for HDF cell migration at P6. Data were analyzed using dependent t-test, One-way ANOVA, or Kruskal-Wallis test.
RESULTS
There were no significant differences in bFGF and KGF mRNA relative expression and HDF migration velocity between the intervention and control groups (p>0.05 and p>0.05, respectively). The doubling time of the KOSR 5% group showed no significant difference (p>0.05), but KOSR 10% and FBS 10% showed significant differences between treatment days 2-6 and treatment days 6-10 (p<0.05).
CONCLUSIONS
The KOSR 10% was comparable to FBS 10% in supporting bFGF and KGF mRNA relative expression, HDF cell proliferation, and HDF cell migration in HDF culture. |
| first_indexed | 2024-03-08T18:26:56Z |
| format | Article |
| id | doaj.art-90211d1aae2949008a6da839e0a263b7 |
| institution | Directory Open Access Journal |
| issn | 2407-2230 1907-3062 |
| language | English |
| last_indexed | 2024-03-08T18:26:56Z |
| publishDate | 2023-09-01 |
| publisher | Faculty of Medicine Trisakti University |
| record_format | Article |
| series | Universa Medicina |
| spelling | doaj.art-90211d1aae2949008a6da839e0a263b72023-12-30T12:04:32ZengFaculty of Medicine Trisakti UniversityUniversa Medicina2407-22301907-30622023-09-01423283293https://doi.org/10.18051/UnivMed.2023.v42.283-293Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblastsNur Ilham Risma Hidayati0https://orcid.org/0009-0001-1310-3151Indra Kusuma1https://orcid.org/0000-0002-7689-451XEko Poerwanto2https://orcid.org/0009-0007-1613-2349Biomedical Science Masters Study Program, Postgraduate School, YARSI University, Jakarta, IndonesiaPhysiology Department, Faculty of Medicine, YARSI University, Jakarta, IndonesiaPhysiology Department, Faculty of Medicine, YARSI University, Jakarta, IndonesiaBACKGROUND Human dermal fibroblast (HDF) cultures can be used as a regenerative agent for wound healing. Fetal bovine serum (FBS) as a culture supplement is derived from animals, therefore not being constant in composition, causes variations in research results, thus requiring a substitute such as KnockOut serum replacement (KOSR). This study evaluated the defined KOSR as FBS substitute for HDF culture by measuring the relative expression of basic fibroblast growth factor (bFGF) and keratinocyte growth factor (KGF) messenger RNA (mRNA), HDF cell proliferation, and HDF migration. METHODS Human dermal fibroblast culture was divided into 2 intervention groups receiving KOSR 5% and KOSR 10%, respectively, and a control group receiving FBS 10%. Reverse transcription polymerase chain reaction (RT-PCR) was used for bFGF and KGF mRNA relative expression at the fifth passage (P5). Cell counting kit-8 (CCK-8) reagent was used for the HDF cell proliferation assay at P5 and the scratch assay was used for HDF cell migration at P6. Data were analyzed using dependent t-test, One-way ANOVA, or Kruskal-Wallis test. RESULTS There were no significant differences in bFGF and KGF mRNA relative expression and HDF migration velocity between the intervention and control groups (p>0.05 and p>0.05, respectively). The doubling time of the KOSR 5% group showed no significant difference (p>0.05), but KOSR 10% and FBS 10% showed significant differences between treatment days 2-6 and treatment days 6-10 (p<0.05). CONCLUSIONS The KOSR 10% was comparable to FBS 10% in supporting bFGF and KGF mRNA relative expression, HDF cell proliferation, and HDF cell migration in HDF culture.https://univmed.org/ejurnal/index.php/medicina/article/view/1476humandermalfibroblastfbskosrbfgfkgfmrnarelativeexpression |
| spellingShingle | Nur Ilham Risma Hidayati Indra Kusuma Eko Poerwanto Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts Universa Medicina humandermalfibroblast fbs kosr bfgf kgf mrnarelativeexpression |
| title | Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| title_full | Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| title_fullStr | Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| title_full_unstemmed | Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| title_short | Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| title_sort | comparative analysis of knockout serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts |
| topic | humandermalfibroblast fbs kosr bfgf kgf mrnarelativeexpression |
| url | https://univmed.org/ejurnal/index.php/medicina/article/view/1476 |
| work_keys_str_mv | AT nurilhamrismahidayati comparativeanalysisofknockoutserumreplacementandfetalbovineserumforinvitrocultureofhumandermalfibroblasts AT indrakusuma comparativeanalysisofknockoutserumreplacementandfetalbovineserumforinvitrocultureofhumandermalfibroblasts AT ekopoerwanto comparativeanalysisofknockoutserumreplacementandfetalbovineserumforinvitrocultureofhumandermalfibroblasts |