Scientists at Cedars-Sinai Medical Center (CSMC) have developed a new method to facilitate the creation of corneal limbal epithelial stem cells (LESC) for patients with limbal epithelial stem cell deficiency (LSCD). LSCD is a condition where the epithelial stem cells in the human cornea degenerate and/or die. LSCD is a fairly common and clinically important cause of corneal blindness. LSCD may develop as a consequence of congenital aniridia, an acquired inflammatory condition or infections, ocular burns, or even long-term contact lens wear.
One approach to treating LSCD is to transplant corneal limbal epithelial stem cells that have been cultured in the lab using human amniotic membrane as a surface on which the cells can grow. Before the amniotic membrance can be used, the amniotic epithelial cells must be removed (a process known as decellularizing) from the human amniotic membrane. Decellularized human amniotic membrane provides a more favorable environment for LESC growth and results in LESCs that are more suitable for transplant. However, most methods to decellularize the membrane are long, tedious, cause tissue damage, and not very reproducible.
Mehrnoosh Saghizadeh, Alex Ljubimov and colleagues have developed a new method for removing live cells from amniotic membrane, described in the November 13, 2013 issue of PLOS One. The method uses the application of an inorganic alkaline solution (sodium hydroxide; NaOH) to the amniotic epithelial cells. Rubbing a mild solution of NaOH on the human amniotic membrane resulted in a visible removal of most amniotic epithelial cells, while leaving major human amniotic membrane components intact. Compared to existing methods, applying NaOH to the amniotic epithelial cells is a fast, easy, and reproducible way to decellularize human amniotic membrane.
Human amniotic membrane is available from placenta discarded following normal deliveries. Dr. Alexander Ljubimov and his multidisciplinary research team made use of a unique service offered by the Clinical and Translational Research Center at CSMC, in which de-identified placentas are collected from the delivery room so that amniotic membrane, segments of umbilical cord, umbilical blood samples, or placental tissue can be used provided to investigators in a variety of research activities. This service is being used by a variety of CTSI investigators and is open to research groups at any of the partner institutions.
PLOS One: A Simple Alkaline Method for Decellularizing Human Amniotic Membrane for Cell Culture