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J. Oh, M. Jung, K. Shin, S. Jeon, M. Kim, J. Lee, W. Wee; Induction of Transdifferentiation of Bone Marrow Stem Cells Into Corneal Epithelial Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):3026.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the possibility to induce the transdifferentiation of bone marrow(BM) stem cells into corneal epithelial cells
Human BM cells were divided into fast adhesion cells(FA) and slow adhesion cells(SA) with DMEM/F12 (3:1) including 10% FBS, 30ng/ml choleratoxin, 2nM 3.3.5 triiodo L–thyronine sodium salt, 0.18mM adenine, 0.4ug/ml hydrocortisone, 5ug/ml insulin, and 5ng/ml EGF for 4 weeks. No feeder was used in group 1, NIH3T3(ATCC:CRL1658) and 3T3–J2 (kindly provided by H. Green) were used as feeder in group 2 and 3. 20ng/ml of FGF was added to cells in the media above mentioned for the 1st week in group 4. Human limbal epithelial cells(HLEC) were cultured in the same media with NIH3T3 feeder as a control. Connexin43(Conx43), cytokeratin12(Ck12), ABCG2, P63, and Oct4 were evaluated using RT–PCR and immunohistochemistry at 2 and 4 weeks. Semiquantitative analysis was done in a ratio of density of the markers to the density of GAPDH with image analyzer(Vilberlourmat) and densitometry(Tina 2.0, Raytest, Straubenhardt, Germany).
The expression of P63, Ck12, and Oct4 was markedly lower in all groups of BM cells compared to HLEC for 4 weeks. The expression of Conx43 increased by 16.7% in group 2 and 58.4% in group 3, and the expression of Oct4 decreased by 66.7% in group 2 and 68.5% in group 3 compared to group 1 at 4 weeks. FGF didn’t induce any change till 2 weeks. There were no differences in the markers mentioned above between FA and SA, irrespective of the media condition or feeders.
Transdifferentiation of BM cells into corneal epithelial cells was not achieved till 4 weeks, but the increase in the expression of Conx43 suggested the possibility of epithelial transdifferentiation using 3T3–J2 as a feeder.
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