Clinical transplantation ofex vivoexpanded autologous limbal epithelial cells using a culture medium with human serum as single supplement: a retrospective case series

Abstract

. Purpose: Presently, our clinic is the only centre in Scandinavia that offers patients with corneal surface pathology including limbal stem cell deficiency (LSCD) transplantation of ex vivo expanded limbal epithelial cells (LECs). We here present clinical data of the first nine patients with LSCD who were transplanted with autologous LECs expanded in medium completely free of any animal‐derived products and non‐human/recombinant growth factors (including Cholera Toxin), and with autologous human serum as the only growth supplement. Methods: We conducted a noncomparative retrospective study of patients with LSCD at our centre between 2009 and 2011. The diagnosis was based on history and clinical signs. A biopsy was taken from healthy limbus, and the epithelium was expanded on amniotic membrane (AM) in medium containing autologous serum and subsequently transplanted to the affected eye. Results: Successful outcome was defined as relief of pain and photophobia and/or improved best corrected visual acuity (BCVA) and/or reestablishment of a stable corneal epithelium and regression of corneal vascularization. Five of the nine transplanted patients (55.6%) had an improvement in either subjective symptoms or objective findings (11‐ to 28‐month follow‐up). Conclusions: Our clinical study shows that patients with LSCD can be treated successfully with transplantation of LECs expanded ex vivo in a medium with autologous serum as the only growth supplement. The use of this novel culture system, which is devoid of animal‐derived products and non‐human/recombinant growth factors (including Cholera Toxin), reduces the risks of inter‐species disease transmission and host immune responses to xenogenic proteins, both obvious advantages for the patient.

Systematic Review and Meta-analysis on Transplantation of Ex Vivo Cultivated Limbal Epithelial Stem Cell on Amniotic Membrane in Limbal Stem Cell Deficiency

“…43 The use of ex vivo cultured LESCs to treat corneal LSCD in humans was first described by Pellegrini et al 44 in 1997. Subsequently, plenty of case series and reviews have reported the efficiency and evidence of this treatment. 12,[25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42][43][45][46][47] However, with the lack of sufficient cases or valid measurements, they failed to provide a convincing assessment of this surgery. Moreover, there is controversy regarding the clinical outcome between different sources of limbal stem cells, such as autograft and allograft.…”

“…may be used as a cell substrate in the ex vivo culture. But human amniotic membrane (HAM), which promotes epithelialization, inhibits fibrosis, has anti-inflammatory and antiangiogenic properties, and possesses antimicrobial and antiviral properties and a high hydraulic conductivity, 22,23 with low or no immunogenicity, 24 has become the widely used material [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40][41][42] acting as a surrogate environmental stem cell niche. 43 The use of ex vivo cultured LESCs to treat corneal LSCD in humans was first described by Pellegrini et al 44 in 1997. Subsequently, plenty of case series and reviews have reported the efficiency and evidence of this treatment.…”

Trypsin for Dissociation of Limbal Cells for Engineering of Grafts May Induce DNA Strand Breaks in the Harvested Cells

“…Here the limbal samples are positioned in medium on a suitable substrate without initial dissociation of the epithelial cells. In recent studies, using this approach, we have demonstrated that transplantable grafts may be generated in a culture system without animal feeder cells and FBS using autologous serum as the single growth promoting supplement [18,19].…”

Human Conjunctival Epithelial Sheets Grown on Poly(Lactic-Co-Glycolic) Acid Membranes and Cocultured With Human Tenon's Fibroblasts for Corneal Repair

“…Moreover, xenobiotic-free cultures are developed using a feeder-free system and either serum-free or human serum-supplemented media to comply with rigorous regulatory standards. 17,18 Most previous research studies have reported on the clinical application of animal cells or animal products, and only three studies have used animal source-free culture systems. 16 Thus, optimization of the culture procedure and the procedural suitability for transplantation are the main concerns when culturing primary hCjECs without using murine feeder layers.…”