The Successful Culture and Autologous Transplantation of Rabbit Oral Mucosal Epithelial Cells on Amniotic Membrane

Nakamura, Takahiro; Endo, Kenichi; Cooper, Leanne J.; Fullwood, Nigel J.; Tanifuji, N; Tsuzuki, Masato; Koizumi, Noriko; Inatomi, Tsutomu; Sano, Yoichiro; Kinoshita, Shigeru

doi:10.1167/iovs.02-0195

Cited by 257 publications

(229 citation statements)

References 36 publications

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“…23,24 As the cultures were not air-lifted, epithelial stratification was less then reported earlier, 25,26 and the cells appeared generally flatter. Interestingly, transcription factor p63, 11 and notably low affinity NGF receptor p75, 27 the progenitor cell markers known to be expressed by oral mucosal epithelial cells were both found in the cultures, along with the expression of ABCG2.…”

Section: Discussionmentioning

confidence: 77%

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Kuo

et al. 2009

Eye

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Purpose To access the feasibility of using cultivated oral mucosal epithelial cell transplantation (COMET) for the management of severe corneal burn. Methods COMET was performed to promote re-epithelialization in two eyes with acute alkaline burn and one eye with chronic alkaline burn, and to reconstruct the ocular surface in two eyes with chronic thermal burn. Autologous oral mucosal epithelial cells obtained from biopsy were cultivated on amniotic membrane. Immunoconfocal microscopy for keratins and progenitor cell markers was performed to characterize the cultivated epithelial sheet. Following transplantation, the clinical outcome and possible complications were documented. The patients were followed for an averaged 29.6 ± 3.6 (range: 26-34) months. Results Cultivated oral mucosal epithelial sheet expressed keratin 3, 13, and progenitor cell markers p63, p75, and ABCG2. After COMET, all the corneas became less inflamed, and the corneal surface was completely reepithelialized in 6.0 ± 3.2 (range: 3-10) days in all but one patients. Microperforation occurred in one patient, and a small persistent epithelial defect developed in another. Both were solved uneventfully. In all patients, superficial corneal blood vessels invariably developed, and to further improve vision, conjunctivo-limbal autografting (N ¼ 3) and/or penetrating keratoplasty (N ¼ 3) were performed subsequently. The vision of all patients showed substantial improvement after additional surgeries. Conclusions This study showed the potential of COMET to promote re-epithelialization and reduce inflammation in acute corneal burn, and to reconstruct the corneal surface in chronic burn. COMET may, therefore, be considered an alternative treatment for severe corneal burn.

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Section: Discussionmentioning

confidence: 77%

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Kuo

et al. 2009

Eye

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“…used in a variety of in vitro and in vivo experiments investigating both corneal pathophysiology (Becker 1961, Starka et al 1977, Green et al 1981, Usukura et al 1988, Felche & Sigler 2002, Nakamura et al 2003, Tungsiripat et al 2003, Ivarsen et al 2004, Mimura et al 2005 and IOP (Becker 1961, Starka et al 1977, Green et al 1981, Usukura et al 1988, Felche & Sigler 2002, Nakamura et al 2003, Tungsiripat et al 2003, Ivarsen et al 2004, Mimura et al 2005. Furthermore, our in-house generated polyclonal antibody to human 11b-HSD1 shared a 93% homology to the rabbit 11b-HSD1 amino acid sequence and the corticosteroid hormone profile is similar to humans (i.e.…”

Section: Western Blot Analysesmentioning

confidence: 89%

“…A possible animal model for these experiments appears to be the New Zealand white albino rabbit (NZWAR). This is an established model for the investigation of corneal disease (Nakamura et al 2003, Tungsiripat et al 2003, Ivarsen et al 2004, Mimura et al 2005 and AH dynamics (Tsukahara et al 1986, Green et al 1989, Munden & Schmidt 1992, Lim et al 2005. The corticosteroid hormone profile is similar to humans (i.e.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of the prereceptor regulation of glucocorticoids in the anterior segment of the rabbit eye

Onyimba¹,

Vijapurapu²,

Curnow³

et al. 2006

Journal of Endocrinology

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The prereceptor regulation of glucocorticoids (GCs) by 11b-hydroxysteroid dehydrogenase type-1 (11b-HSD1), a bidirectional isozyme that interconverts active (cortisol) and inactive (cortisone) GCs, is an established determinant of GC function in tissues such as liver, adipose and bone. Although the therapeutic use of GCs is abundant in ophthalmic practice, where GC interactions with nuclear receptors modulate gene transcription, the prereceptor regulation of endogenous cortisol is not well described in ocular tissues. Recent descriptive studies have localised 11b-HSD1 to the human corneal epithelium and non-pigmented epithelium (NPE) of the ciliary body, indicating a link to corneal epithelial physiology and aqueous humour production. In this study, we characterise the functional aspects of the autocrine regulation of GCs in the anterior segment of the rabbit eye. Using our in-house generated primary antibody to human 11b-HSD1, immunohistochemical analyses were performed on paraffin-embedded sections of whole New Zealand white albino rabbits, (NZWAR) eyes. As in human studies, 11b-HSD1 was localised to the corneal epithelium and the NPE. No staining was seen in the albino 'pigmented' ciliary epithelium. Specific enzyme assays for oxo-reductase (cortisone/cortisol) and dehydrogenase (cortisol/cortisone) activity indicated predominant 11b-HSD1 oxo-reductase activity from both the intact ciliary body tissue (nZ12, median 2$1 pmol/mg per h and range 1$25-2$8 pmol/mg per h; PZ0$006) and primary cultures of corneal epithelial cells (nZ12, median 3$0 pmol/mg per h and range 1$0-7$4 pmol/mg per h, PZ0$008) compared with dehydrogenase activity (median 1$0 pmol/mg per h and range 0$5-2$0 pmol/mg per h; median 0$5 pmol/mg per h and range 0$25-1$9 pmol/mg per h respectively). These findings were supported by expression of 11b-HSD1 protein as visualised by Western blotting of ciliary body tissue and immunocytochemistry of corneal epithelial cells. Reduction of corneal epithelial cell proliferation was seen after primary cultures were co-incubated with cortisol and cortisone. 11b-HSD1 activity was not demonstrated in naïve conjunctival fibroblasts or corneal stromal keratocytes. Our results indicate that the distribution of 11b-HSD1 in the rabbit resembles that of the human eye and activates cortisone to cortisol in both corneal and uveal tissues. The NZWAR provides a suitable in vivo model for the further evaluation of 11b-HSD1 activity in the eye, especially its role in corneal epithelial and ciliary body physiology.

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“…Therefore, we focused on epithelial stem cells derived from oral mucosa as an alternative to limbal epithelial cells used in the autologous regenerative therapy of ocular surfaces (Figure 4). In fact, bioengineered tissues derived from oral mucosal epithelial cells showed similar optical transparency compared to the corneal epithelium and were available for transplantation to a rabbit corneal model [55]. Moreover, the substitution of limbal epithelial cells by oral mucosal epithelial cells is also supported by the histological observation that human oral mucosal cell sheets recapitulated a structure comparable to that of the native corneal epithelium [36].…”

Section: Reconstruction Of the Corneal Epitheliummentioning

confidence: 97%

Regenerative medicine of cornea by cell sheet engineering using temperature-responsive culture surfaces

et al. 2013

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Recently, regenerative medicine has been focused on as next-generation definitive therapies for several diseases or injuries for which there are currently no effective treatments. These therapies have been rapidly developed through the effective fusion between different fields such as stem cell biology and biomaterials. So far, we have proposed "cell sheet engineering" through our core technology that simply applies alterations of the temperature which allows regulation of the attachment or detachment of living cells on the culture surfaces grafted with the temperature-responsive polymer "poly(N-isoproplyacrylamide)". This technology enables us to construct bioengineered sheet-like tissues, termed "cell sheets", without the need of using biodegradable scaffolds and protease treatments that are traditionally used. Therefore, our carrier-free cell sheets not only are independent of the issues observed in conventional methods, but also showed further advantages in the reconstruction of the corneal epithelium or endothelium (e.g. improvement of optical transparency and cell-cell interactions to host stroma in reconstructed tissues). Moreover, our approach does not have issues such as immune rejection or limited number of donors, due to the use of cell sheets derived from autologous limbal (in patients with unilateral disease) or oral mucosal epithelial cells (in patients with bilateral disorders). Indeed, we have successfully performed the clinical application for the reconstruction of ocular surfaces through the transplantation of our carrier-free corneal epithelial cell sheets, as evidenced by improved visual acuity as well as long-term maintenance of postoperative health conditions on ocular surfaces in all patients. We have also proposed a novel approach for the reconstruction of the corneal endothelium using corneal endothelial cell sheets by demonstrating its successful application. Thus, our cell sheet engineering technique provides a breakthrough in the field of regenerative medicine applied to the cornea. cell sheet, temperature-responsive, carrier-free Citation:Umemoto T, Yamato M, Nishida K, et al. Regenerative medicine of cornea by cell sheet engineering using temperature-responsive culture surfaces. Chin Sci Bull, 2013, 58: 43494356,

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The Successful Culture and Autologous Transplantation of Rabbit Oral Mucosal Epithelial Cells on Amniotic Membrane

Cited by 257 publications

References 36 publications

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Transplantation of cultivated oral mucosal epithelial cells for severe corneal burn

Characterisation of the prereceptor regulation of glucocorticoids in the anterior segment of the rabbit eye

Regenerative medicine of cornea by cell sheet engineering using temperature-responsive culture surfaces

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