Human serum is an advantageous supplement for human dermal fibroblast expansion: clinical implications for tissue engineering of skin.
BACKGROUND
Standard fibroblast culture medium usually contains fetal bovine serum (FBS). In theory, unknown risks of infection from bovine disease or immune reaction to foreign proteins may occur if standard culture method is used for future human tissue-engineering development. Human serum (HS) theoretically would be another choice in providing a safer approach and autologous clinically reliable cells.


METHODS
Isolated human dermal fibroblasts were culture-expanded in an equal volume mixture of Ham's F12 medium and Dulbecco's Modified Eagle Medium (DMEM) supplemented with either 10% HS or 10% FBS from passage 0 to passage 3. Effects of 10% HS and 10% FBS on human fibroblast viability, growth kinetics, cell cycle analysis and gene expressions were investigated and compared.


RESULTS
Generally, fibroblast viability cultured in HS supplementations was much higher compared to FBS supplementation. Fibroblast proliferations were faster in HS supplementations with shorter doubling time. Cell cycle analysis showed fibroblasts cultured with HS supplementations have higher S-phase ratio compared to FBS. Gene expression levels by quantitative reverse transcriptase-polymerose chain reaction (RT-PCR) showed cultured fibroblasts with HS supplementation maintains expression of collagen type I collagen, increased expression of type III collagen and fibronectin and reduced expression of alpha-smooth muscle actin (alpha-SMA) compared to FBS.


CONCLUSIONS
Results demonstrated potential advantages of HS vs. FBS in generating larger numbers of cultured dermal fibroblasts in a shorter period of time. HS also influenced mRNA expression of type III collagen and fibronectin (upregulated) and alpha-SMA (downregulated), which are important extracellular matrix proteins in wound healing.
Mazlyzam
Abdul Latif
AL
Department of Biomedical Sciences, Faculty of Allied Health Sciences, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.
Aminuddin
Bin Saim
BS
Saim
Lokman
L
Ruszymah
Binti Haji Idrus
BH
D000328:Adult; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D002453:Cell Cycle / Q000502:physiology; D048430:Cell Shape; D002470:Cell Survival; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D044967:Serum / Q000378:metabolism*; D012867:Skin / Q000166:cytology*; D023822:Tissue Engineering