Effect of supplementation of dermal fibroblasts conditioned medium on expansion of keratinocytes through enhancing attachment.
In the present study in vitro expansion of human keratinocytes by supplementing dermal fibroblasts conditioned medium (DFCM) has been reported. Effect of two different DFCM acquired by culturing fibroblasts in keratinocyte-specific medium (defined keratinocytes serum free medium, DFCM-DKSFM) and fibroblast-specific serum free medium (F12: DMEM nutrient mix, DFCM-FD) have been compared. Growth kinetics of keratinocytes in terms of efficiency of cell attachment, expansion index, apparent specific growth rate and growth potential at the end of culture was evaluated in culture supplemented with DFCM-DKSFM and DFCM-FD in comparison with control i.e. DKSFM only. Results indicated that supplementation of DFCM caused significant increase in keratinocyte attachment. Efficiency of keratinocyte attachment in culture supplemented with bFCM-DKSFM was significantly higher compared to those cultured in DFCM-FD and DKSFM. In addition, the expansion index of keratinocytes in cultures supplemented with DFCM-DKSFM and DFCM-FD were 3.7 and 2.2 times higher than that of control condition even though the apparent growth rate and proliferative potential was found significantly lower. These results suggested that supplementation of DFCM enhanced expansion of keratinocyte by increasing efficiency of cell attachment, and DFCM-DKSFM provided suitable condition for in vitro expansion of keratinocytes compared to DFCM-FD and control condition.
Tissue Engineering Centre, Universiti Kebangsaan Malaysia (National University of Malaysia) Medical Centre Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia.
S R
SR
Chowdhury
B S
BS
Aminuddin
B H I
BH
Ruszymah
D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D016895:Culture Media, Serum-Free; D020405:Dermis / Q000166:cytology; D005260:Female; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*; D008875:Middle Aged
D017077:Culture Media, Conditioned; D016895:Culture Media, Serum-Free