The optimum oxygen level in hypoxic culture conditions of ligament derived stem cells: experimental research
Penulis/Author
dr. Sholahudin Rhatomy, Sp.BO(K) (1); Prof. Dr. Dwikora Novembri Utomo, dr., Sp.OT(K) (2); Prof. Dr. dr. Cita Rosita Sigit Prakoeswa, Sp.KK(K) (3); Dr. Heri Suroto, dr.,SpOT(K) (4); Damayanti Tinduh (5); Hari Basuki Notobroto (6); dr. Nur Arfian, Ph.D. (7); Prof. Dr. drh. Fedik Abdul Rantam (8); Ferdiansyah Mahyudin (9)
Tanggal/Date
23 2023
Kata Kunci/Keyword
Abstrak/Abstract
The hypoxic condition is a physiological norm for various stem cells. The natural microenvironment contains lower oxygen pressures. Recent studies reported significant increases in the cultured cells' proliferation in the presence of a low oxygen pressure.
Objective: This study aimed to investigate the optimum oxygen level for rabbit cruciate ligament fibroblast cells culture and Ligament Derived Conditioned Medium/LD-CM (Secretome) preparation in vitro.
Materials and methods: Fibroblasts were isolated from the cruciate ligament of the rabbit's knee. Cultured of rabbit cruciate ligament Fibroblast Cells (fifth passage) were assigned to the slight (5% O2), middle (3% O2), and severe hypoxia (1% O2) groups and the normoxia (21% O2) group. Measurement of growth factors: TGF-β1, PDGF, FGF, and VEGF in LD-CM (Secretome) used an enzyme-linked immunosorbent assay.
Results: The highest number of cultured cells were in the 5% O2 group compared to the normoxia, 1 and 3% groups. The hypoxia 5% group also had increased productions of PDGF, FGF, and VEGF proteins in LD-CM (secretome) compared to the 1, 3%, and normoxia groups. TGF-β1 production was slightly higher in the 3 group than the 5% group.
Conclusion: The hypoxic precondition of 5% oxygen was the optimum condition for ligament culture and ligament derived conditioned medium (secretome) preparation in vitro.
Keywords: hypoxia; ligament derived conditioned medium; normoxia; proliferation; secretome
