A preliminary study of mesenchymal stem cell-like cells derived from murine corneal stroma.
BACKGROUND
Mesenchymal stem cells can be isolated from various tissues besides bone marrow and can differentiate into cells of three germ layers. Recent studies indicate that some cells in corneal stroma express stem cell markers and can also differentiate into chondrocytes and neurocytes. This study was carried out to investigate whether mesenchymal stem cells reside in the murine corneal stroma.


METHODS
Corneas of BALB/c mice were treated with collagenase digestion after the epithelium and endothelium were removed. Then the single cells were harvested and further identified by reverse transcription polymerase chain reaction (RT-PCR). After the immunophenotype of passage 2 corneal stroma-derived cells was analyzed by flow cytometry, attempts were made to differentiate these cells into adipocytes and osteocytes using conditioned medium. Following induction, cells were evaluated by RT-PCR, oil red O and Alizarin Red staining.


RESULTS
Isolated single cells were of stromal origin, not of epithelial or endothelial. Passage 2 corneal stroma-derived cells exhibited the spindle-shaped morphology and expressed CD29, CD90, CD105, and CD71; but were negative for CD34 and CD45. In addition, these cells showed the potentiality of differentiating into adipocytes and osteocytes, which was confirmed by RT-PCR and staining.


CONCLUSIONS
This study demonstrates the presence of mesenchymal stem cell-like cells in the murine corneal stroma. Further analysis of these cells will aid elucidation of the mechanisms of some keratopathies, and these cells may be a source for bioengineering of corneal tissue and for cell-based therapeutics.
Lu
Jian-Min
JM
Department of Ophthalmology, Third Hospital of Hebei Medical University, Ziqiang Road No. 139, Shijiazhuang, China.
Zhou
Zhong-You
ZY
Zhang
Xiao-Rong
XR
Li
Xiao-Lei
XL
Wang
Hui-Fang
HF
Song
Xiu-Jun
XJ
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D003319:Corneal Stroma / Q000166:cytology*; D005434:Flow Cytometry; D016130:Immunophenotyping; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D020133:Reverse Transcriptase Polymerase Chain Reaction
D015703:Antigens, CD; D015415:Biomarkers
Invest Ophthalmol Vis Sci. 2007 Sep;48(9):4061-9
17724188
Int J Hematol. 2005 Feb;81(2):126-30
15765780
Stem Cells. 2006 Dec;24(12):2714-22
16888282
Mol Vis. 2009;15:99-107
19156227
FASEB J. 2005 Aug;19(10):1371-3
15901670
Mol Vis. 2008 Jan 30;14:197-203
18334932
Invest Ophthalmol Vis Sci. 2007 Nov;48(11):5038-45
17962455
Exp Eye Res. 2009 Aug;89(2):209-17
19328783
Cytotherapy. 2007;9(3):252-8
17464757
J Cell Biochem. 2009 Apr 15;106(6):984-91
19229871
Stem Cells. 2005 Apr;23(4):507-15
15790772
Braz J Med Biol Res. 2003 Aug;36(8):1037-46
12886457
Exp Eye Res. 2005 Sep;81(3):247-64
16051216
Stem Cells. 2009 Jul;27(7):1635-42
19544455
Clin Exp Ophthalmol. 2003 Apr;31(2):147-54
12648050
Blood. 2003 Aug 15;102(4):1548-9
12900350
Histochem Cell Biol. 2007 Dec;128(6):551-5
17851677
Mol Vis. 2008 Mar 04;14:431-42
18334960
Exp Hematol. 2000 Aug;28(8):875-84
10989188
Cloning Stem Cells. 2004;6(4):369-74
15671665
Graefes Arch Clin Exp Ophthalmol. 1998 Jul;236(7):522-6
9672798
Stem Cells. 2005 Oct;23(9):1266-75
16051989
J Cell Biochem. 2006 Dec 1;99(5):1285-97
16795045
Invest Ophthalmol Vis Sci. 2006 Feb;47(2):545-51
16431949
Invest Ophthalmol Vis Sci. 2003 Feb;44(2):548-57
12556382
J Cell Sci. 2006 Jun 1;119(Pt 11):2204-13
16684817
Arch Ophthalmol. 2006 Jan;124(1):62-9
16401786
Invest Ophthalmol Vis Sci. 2006 May;47(5):1918-27
16638999
Invest Ophthalmol Vis Sci. 1999 Jul;40(8):1658-63
10393032
Invest Ophthalmol Vis Sci. 2002 Nov;43(11):3416-21
12407151