Abstract
Purpose:
Poor connection and adhesion of epithelial cells to underlying connective tissues is the basic defect of blister disease of conjunctiva, skin, and mouth mucosal membrane such as ocular cicatricial pemphigoid. Transplantation of extracellular matrix (ECM) along with the autologous buccal mucosal epithelial cells may modulate the survival and subsequent proliferation of transplanted epithelial cells. Thus, we have developed a technique to transfer native ECM with human mucosal epithelial cells in vitro.
Methods:
Confluent monolayer human mucosal epithelial cells in the culture plate was pretreated with 0.25% edetic acid for 15 minutes and coated with a 800 μ layer of 20% gelatin. Patches of the mucosal epithelial cells were harvested and transferred to another culture plate. Cell viability and the ability of the transferred cells to proliferate in culture were determined. The ECM transferred along with the cells was characterized by immunohistochemistry.
Results:
Human mucosal epithelial cells in culture can be harvested as an organized cell patch with high efficiency (92.3 ± 4.5%) and high cell viability (89.5 ± 5.5%). Cells harvested from tissue culture plates divided and became confluent within 21 days. Immunohistochemistry demonstrates that the transferred ECM along contains laminin and type IV collagen.
Conclusions:
We were able to harvest mucosal epithelial cells as an organized monolayer from tissue culture plate along with their intercellular extracellular matrix. This approach may provide a practical technique for isolating and transplanting cultivated autologous epithelial cells to reconstruct the damaged ocular surface in Steven-Johnson syndrome, chemical and thermal injury, and ocular cicatricial pemphigoid.
Keywords: 482 cornea: epithelium •
741 transplantation •
519 extracellular matrix