First Autologous Transplantation of Ex Vivo Expanded Limbal Epithelial Cells on Amniotic Membrane Using Air-Liquid Interface
The first autologous transplantation of ex vivo expanded human limbal epithelial cells cultured on acellular human amniotic membrane using air-liquid interface for ocular surface reconstruction was performed in 2006. 1
Historical Development of the Technique
The technique described in 2006 by Sangwan et al. in the Journal of Postgraduate Medicine represented a significant advancement in ocular surface reconstruction. This simple feeder-cell free technique involved:
- Harvesting small limbal tissues (2 mm) from patients with limbal stem cell deficiency (LSCD)
- Proliferating these tissues in vitro on human amniotic membrane
- Supplementing with human corneal epithelial cell medium and autologous serum
- Creating an air-liquid interface culture system
- Cultivating until covering ≥50% of the 2.5 x 5 cm amniotic membrane area
The 2006 publication documented 542 cultures from 250 limbal tissues cultivated between January 2001 and July 2005, with successful explant cultures observed in 98.5% of cases 1.
Evolution of the Technique
Following this initial development, the technique has evolved with several refinements:
In 2010, this approach was successfully used to treat a case of partial LSCD caused by epidermolysis bullosa dystrophica, demonstrating its clinical efficacy in complex cases 2
By 2012, researchers had modified the protocol to use human serum as a single growth supplement, eliminating the need for animal-derived products like fetal bovine serum, cholera toxins, and exogenous growth factors 3
In 2013, a novel method of cocultivating autologous limbal and conjunctival epithelium on a single amniotic membrane substrate was developed for severe cases involving both limbal stem cell deficiency and conjunctival damage 4
By 2016, a standardized protocol was established that could uniformly promote growth and differentiation of cells from limbal, conjunctival, or oral mucosal biopsies into corneal lineage using recombinant epidermal growth factor and insulin 5
Clinical Significance
This technique represented a breakthrough in "cell therapy" for reconstructing the ocular surface in severe limbal stem cell deficiency, particularly for cases caused by chemical burns. The approach offers several advantages:
- Requires only a small limbal biopsy (2 mm) rather than direct limbal tissue transplantation
- Produces a complete monolayer of epithelial cells within 10-14 days
- Achieves successful culture in over 98% of attempts
- Allows for clinical transplantation after just 10-14 days of cultivation
Current Applications
Today, corneal transplantation, including techniques using cultivated limbal epithelial cells, remains the most common therapeutic option for patients with corneal damage and reduced vision or significant pain due to bullous keratopathy 6.
The understanding of preserving stem cells has led to widespread use of amniotic membrane in various techniques, including "inlay" methods where the membrane acts as a scaffold for epithelial cell migration, and "overlay" methods where it functions as a biological contact lens 6.