The role of signaling pathways in the expansion of corneal epithelial cells in serum-free B27 supplemented medium.
OBJECTIVE
To study the influence of serum-free B27 supplemented culture medium on corneal epithelial cells from limbal explants.


METHODS
Human limbal tissues obtained from cadaveric donor eyes were used in this study. The morphological characteristics of cultivated epithelial cells were analyzed by phase contrast microscopy. Growth kinetics, bromodeoxyuridine (BrdU) labeling cell proliferation assay, and reverse transcriptase PCR (RT-PCR) for limbus and corneal markers were studied in serum-dependent and serum-free B27 supplemented corneal epithelial culture. The signaling pathway genes were analyzed by RT(2) qPCR profiler array.


RESULTS
The corneal epithelial cells morphology and mRNA expression of markers were similar in both the serum-dependent and serum-free B27 supplemented culture. The growth and proliferation of the serum-free B27 supplemented culture was significantly higher than that of the serum-dependent culture. The wnt, hedgehog, survival, NFkB, Jak-Stat, and calcium protein kinase C pathways were highly expressed in the serum-free B27 supplemented corneal epithelial culture.


CONCLUSIONS
Most signaling pathway genes are upfolded by B27 supplementation in the corneal epithelial cell culture; it could be an efficient replacement for serum.
Krishnan
Sasirekha
S
L&T Department of Ocular Pathology, Vision Research Foundation, Sankara Nethralaya, Chennai, India. krishnan.sasirekha@gmail.com
Lakshmanan
Shruthi
S
Iyer
Geetha Krishnan
GK
UmaMaheswari
Krishnan
K
Krishnakumar
Subramanian
S
D000368:Aged; D000369:Aged, 80 and over; D001973:Bromodeoxyuridine / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D019573:Epithelium, Corneal / Q000166:cytology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects* / Q000235:genetics
D016895:Culture Media, Serum-Free; D012333:RNA, Messenger; D001973:Bromodeoxyuridine