Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells

Osei-Bempong, Charles; Ghareeb, Ali E.; Lako, Majlinda; Figueiredo, Francisco C; Armitage, WJ

doi:10.1016/j.cryobiol.2018.07.008

Cited by 14 publications

(8 citation statements)

References 15 publications

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“…With the development of cryopreservation medical technology, the technology of cryopreservation of cells and single tissue has been quite mature in recent studies [16]. However, the preservation of more complex large-sized composite tissues is still in the experimental exploration stage.…”

Section: Discussionmentioning

confidence: 99%

Dimethyl Sulfoxide is Less Effective in Immersing Cryopreserved Large Pieces of Tissue: A Rabbit Hind-Limb Model

et al. 2019

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BackgroundDimethyl sulfoxide (DMSO) cryoprotectant can effectively alleviate the damage to single tissue during cryopreservation and restore its physiological activity after rewarming. However, studies have not been successful for preserving large tissue. This study aimed to investigate the application conditions of DMSO in large composite-tissue by performing femoral artery perfusion and soaking in a rabbit hind-limb model.Material/MethodsA microdialysis-freezing point osmometer was used to detect the minimum time required for effective perfusion of 10% v/v perfusion and 20% v/v perfusion group. Magnetic resonance spectroscopy (MRS) was used to detect the area under the spectrum peak of DMSO in perivascular, intramuscular, subcutaneous areas, and compare the area under the spectrum peak in the 20% vascular perfusion group and other whole immersion groups.ResultsThe minimum time required for effective perfusion of muscle in the 10% v/v group was 30 minutes, the DMSO concentration was stable at 5% v/v; In the 20% v/v group the minimum time was at 20 minutes, stable at 12% v/v. There was a statistically difference of the area under the spectrum peak in the 10% group and the 20% v/v group after vascular perfusion in different tissue locations (P<0.05). The 20% vascular perfusion group and the different concentration of DMSO in the large tissue soaking group were statistically different (P<0.05). There was a significant difference in the 20% v/v vascular perfusion group compared to the low concentration immersion group, but no difference compared to the 50% immersion group.ConclusionsThe effect of blood perfusion on cryopreservation in large tissue by using DMSO was slightly better than overall soaking, especially in preservation of skin and subcutaneous tissue.

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Section: Discussionmentioning

confidence: 99%

Dimethyl Sulfoxide is Less Effective in Immersing Cryopreserved Large Pieces of Tissue: A Rabbit Hind-Limb Model

et al. 2019

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“…Long-term storage of cultivated LSCs may allow re-grafting in cases of relapse, without the need for further biopsy. Hence, cryopreservation of cultivated LSCs can potentially allow their indefinite storage for re-graft and transport from centralised laboratories [85,96,97].…”

Section: Cultivated Limbal Epithelial Transplant (Clet)mentioning

confidence: 99%

Recent Advances in Stem Cell Therapy for Limbal Stem Cell Deficiency: A Narrative Review

2020

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Destruction of the limbus and depletion of limbal stem cells (LSCs), the adult progenitors of the corneal epithelium, leads to limbal stem cell deficiency (LSCD). LSCD is a rare, progressive ocular surface disorder which results in conjunctivalisation and neovascularisation of the corneal surface. Many strategies have been used in the treatment of LSCD, the common goal of which is to regenerate a self-renewing, transparent, and uniform epithelium on the corneal surface. The development of these techniques has frequently resulted from collaboration between stem cell translational scientists and ophthalmologists. Direct transplantation of autologous or allogeneic limbal tissue from a healthy donor eye is regarded by many as the technique of choice. Expansion of harvested LSCs in vitro allows smaller biopsies to be taken from the donor eye and is considered safer and more acceptable to patients. This technique may be utilised in unilateral cases (autologous) or bilateral cases (living related donor). Recently developed, simple limbal epithelial transplant (SLET) can be performed with equally small biopsies but does not require in vitro cell culture facilities. In the case of bilateral LSCD, where autologous limbal tissue is not available, autologous oral mucosa epithelium can be expanded in vitro and transplanted to the diseased eye. Data on long-term outcomes (over 5 years of follow-up) for many of these procedures is needed, and it remains unclear how they produce a self-renewing epithelium without recreating the vital stem cell niche. Bioengineering techniques offer the ability to recreate the physical characteristics of the stem cell niche, while induced pluripotent stem cells offer an unlimited supply of autologous LSCs. In vivo confocal microscopy and anterior segment OCT will complement impression cytology in the diagnosis, staging, and follow-up of LSCD. In this review we analyse recent advances in the pathology, diagnosis, and treatment of LSCD.

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“…Improving the storage of HLECs is important for providing a reliable source of tissue for treating limbal stem cell deficiency. If the limbal tissue taken from the healthy eye is too big, it risks jeopardising the limbal stem cells in the healthy eye [ 59 ]. So cryopreservation (4°C) has the potential to indefinitely extend the lifespan of LSCs.…”

Section: Application Of Hypothermia In An Eye Bankmentioning

confidence: 99%

Research Progress of the Application of Hypothermia in the Eye

2020

Oxidative Medicine and Cellular Longevity

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Hypothermia is widely used in the medical field to protect organs or tissues from damage. Different research fields have different explanations of the protection mechanism of hypothermia. Hypothermia is also widely used in the field of ophthalmology, for example, in the eye bank, the preservation of corneal tissue and the preservation of the eyeball. Low temperature can also be applied to some ophthalmic diseases, such as allergic conjunctivitis, retinal ischemia, and retinal hypoxia. It is used to relieve eye symptoms or reduce tissue damage. Hypothermic techniques have important applications in ophthalmic surgery, such as corneal refractive surgery, vitrectomy surgery, and ciliary body cryotherapy for end-stage glaucoma. Hypothermia can reduce the inflammation of the cornea and protect the retinal tissue. The eyeball is a complex organ, including collagen tissue of the eyeball wall and retinal nerve tissue and retinal blood vessels. The mechanism of low temperature protecting eye tissue is complicated. It is important to understand the mechanism of hypothermia and its applications in ophthalmology. This review introduces the mechanism of hypothermia and its application in the eye banks, eye diseases (allergic conjunctivitis, retinal ischemia, and hypoxia), and eye surgeries (corneal transplant surgery, corneal refractive surgery, and vitrectomy).

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Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells

Cited by 14 publications

References 15 publications

Dimethyl Sulfoxide is Less Effective in Immersing Cryopreserved Large Pieces of Tissue: A Rabbit Hind-Limb Model

Dimethyl Sulfoxide is Less Effective in Immersing Cryopreserved Large Pieces of Tissue: A Rabbit Hind-Limb Model

Recent Advances in Stem Cell Therapy for Limbal Stem Cell Deficiency: A Narrative Review

Research Progress of the Application of Hypothermia in the Eye

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