J Korean Med Sci.  2014 Jun;29(6):846-851. 10.3346/jkms.2014.29.6.846.

Evaluation of Stem Cell Components in Retrocorneal Membranes

• Lee SH
• Kim KW
• Kim MK
• Chun YS
• Kim JC

• KMID: 1796949
• DOI: http://doi.org/10.3346/jkms.2014.29.6.846

Abstract

The purpose of this study was to elucidate the origin and cellular composition of retrocorneal membranes (RCMs) associated with chemical burns using immunohistochemical staining for primitive cell markers. Six cases of RCMs were collected during penetrating keratoplasty. We examined RCMs with hematoxylin and eosin (H&E), periodic acid-Schiff (PAS) staining and immunohistochemical analysis using monoclonal antibodies against hematopoietic stem cells (CD34, CD133, c-kit), mesenchymal stem cells (beta-1-integrin, TGF-beta, vimentin, hSTRO-1), fibroblasts (FGF-beta, alpha-smooth muscle actin), and corneal endothelial cells (type IV collagen, CD133, VEGF, VEGFR1). Histologic analysis of RCMs revealed an organized assembly of spindle-shaped cells, pigment-laden cells, and thin collagenous matrix structures. RCMs were positive for markers of mesenchymal stem cells including beta-1-integrin, TGF-beta, vimentin, and hSTRO-1. Fibroblast markers were also positive, including FGF-beta and alpha-smooth muscle actin (SMA). In contrast, immunohistochemical staining was negative for hematopoietic stem cell markers including CD34, CD133 and c-kit as well as corneal endothelial cell markers such as type IV collagen, CD133 except VEGF and VEGFR1. Pigment-laden cells did not stain with any antibodies. The results of this study suggest that RCMs consist of a thin collagen matrix and fibroblast-like cells and may be a possible neogenetic structure produced from a lineage of bone marrow-derived mesenchymal stem cells.

Keyword

Burns, Chemical; Cornea; Mesenchymal Stem Cell; Retrocorneal Membrane

MeSH Terms

Adult
Aged
Antigens, CD/metabolism
Cornea/*cytology/pathology
Cytokines/metabolism
Endothelial Cells/cytology/metabolism
Female
Fibroblasts/cytology/metabolism
Hematopoietic Stem Cells/cytology/metabolism
Humans
Immunohistochemistry
Intercellular Signaling Peptides and Proteins/metabolism
Male
Mesenchymal Stromal Cells/cytology/metabolism
Middle Aged
Stem Cells/cytology/*metabolism
Antigens, CD
Cytokines
Intercellular Signaling Peptides and Proteins

Figure

• 

  Fig. 1 Histochemical findings of the retrocorneal membranes. (A) H&E staining of the Case 1 demonstrates avascular connective tissue with spindle-shaped cells (arrow head) and pigment-laden cells (red arrow). (B) PAS staining of the Case 3 reveals no specific findings. (C) Masson's trichrome staining detects a collagen fiber structure within the retrocorneal membranes (Case 3). (B & C) In Case 3, there are also spindle-shaped cells (arrow head) and pigment-laden cells (red arrow) in both of PAS staining and Masson's trichrome staining photos (A, B, C: ×400).

• 

  Fig. 2 Spindle-shaped cells in the retrocorneal membranes were positive for FGF-β (Case 4, A), TGF-β (Case 4, B), α-SMA (Case 4, C), β1-integrin (Case 5, D), vimentin (Case 5, E) and hSTRO-1 (Case 6, F) (arrow heads). Pigment-laden cells do not react with any of the antibodies (red arrows) (A-F: ×400).

• 

  Fig. 3 CD34 (Case 6, A) and c-kit (Case 6, B) are not detected in any cells within the retrocorneal membrane. In contrast, VEGF (Case 4, C) and VEGF receptor 1 (Case 4, D) are slightly expressed (arrow heads). Red arrows indicate pigment-laden cells (A-D: ×400).

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