Validation System of Tissue-Engineered Epithelial Cell Sheets for Corneal Regenerative Medicine

Hayashi, Ryuhei; Yamato, Masayuki; Takayanagi, Hiroshi; Oie, Yoshinori; Kubota, Akira; Hori, Yuichi; Okano, Teruo; Nishida, Kohji

doi:10.1089/ten.tec.2009.0277

Cited by 36 publications

(35 citation statements)

References 21 publications

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“…To analyze the phenotype of the cultured cells, the number of cultured OMEC that expressed p63 and ABCG2 was calculated. Although p63 expression has been observed by immunostaining in several studies on cultured OMEC, 11,14,38,39 to our knowledge, only the study by Oie et al provide quantitative data. 38 The authors compared the effects of two types of irradiated feeder layers (NIH/3T3 and dermal fibroblasts) on cultured OMEC.…”

Section: Discussionmentioning

confidence: 90%

Xenobiotic- and Serum-Free Culture of Oral Mucosal Epithelial Cells on Contact Lenses

Björkblom¹,

Eidet²,

Utheim³

et al. 2015

Current Eye Research

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Section: Discussionmentioning

confidence: 90%

Xenobiotic- and Serum-Free Culture of Oral Mucosal Epithelial Cells on Contact Lenses

Björkblom¹,

Eidet²,

Utheim³

et al. 2015

Current Eye Research

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“…Translation of this antiapoptotic approach toward storage of donor corneas and corneal transplantation could decrease the amount of discarded donor tissue and increase the quality of patient care, as grafts with a higher EC count are less likely to undergo graft failure, resulting in a decreased requirement for regrafting. Last, this approach could be applicable to other forms of cell transplantation procedures, such as use of engineered stem cell sheets (Hayashi et al, 2009) or skeletal myocardial sheet transplantation to improve the damaged heart (Shimizu et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

Corneal Endothelial Cells Are Protected from Apoptosis by Gene Therapy

et al. 2011

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Corneal grafting is the most prevalent form of transplantation. Corneal endothelial cells (ECs), which form a monolayer of the cornea with minimal proliferative potential, are pivotal for maintenance of corneal clarity. Loss of EC viability and apoptosis leads to graft failure posttransplantation and reduces the quality of donor corneas in storage, such that up to 30% do not meet selection criteria and must be discarded. The current study investigates antiapoptotic effects of transduced mammalian Bcl-x(L) and baculoviral p35 on human ECs. Multiple apoptotic cell features are observed while inducing apoptosis either via the extrinsic (death receptor) or intrinsic (mitochondrial) apoptotic pathway. Human ECs were studied under three experimental conditions: (1) as an immortalized cell line, (2) as primary cells, and (3) in an intact cornea. Interestingly, in primary EC suspensions, Bcl-x(L) was protective against apoptosis mediated via both pathways. However, p35 was significantly more protective against apoptosis mediated via the intrinsic pathway compared with Bcl-x(L). Our results provide critical insight into the role of apoptotic pathways in the maintenance of EC viability and the efficacy with which these protective proteins exert their effect. These observations could form the basis for future applications of antiapoptotic gene therapy to corneal preservation aiming to reduce both graft failure after transplantation as well as donor corneal damage during storage.

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“…In fact, our clinical trials using autologous oral mucosal cell sheets prepared using temperature-responsive culture surfaces have demonstrated significant improvement in visual acuity and successful long-term maintenance of healthy conditions at the ocular surface in all patients after transplantation ( Figure 5) [36]. In addition, we recently established more optimized culture conditions in the preparation of cell sheets [57][58][59] and the validation system for tissue-engineered cell sheets [60,61]. Moreover, we also developed an autologous transplantation technique for patients with inherited corneal disorders by genetic modification of corneal epithelial stem cell sheets using lentiviral vectors [62].…”

Section: Reconstruction Of the Corneal Epitheliummentioning

confidence: 99%

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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Validation System of Tissue-Engineered Epithelial Cell Sheets for Corneal Regenerative Medicine

Cited by 36 publications

References 21 publications

Xenobiotic- and Serum-Free Culture of Oral Mucosal Epithelial Cells on Contact Lenses

Xenobiotic- and Serum-Free Culture of Oral Mucosal Epithelial Cells on Contact Lenses

Corneal Endothelial Cells Are Protected from Apoptosis by Gene Therapy

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

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