Microcarrier cell culture of neonatal human corneal endothelium

Insler, Michael S.; Lopez, Jamie G.

doi:10.3109/02713689009000051

Cited by 16 publications

(11 citation statements)

References 11 publications

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“…[3][4][5][6] Previous studies showed that HCECs can be successfully cultured by the stimulation of growth factors. [7][8][9][10][11][12] This means that HCECs do not lose their proliferative capacity, but retain the potential to proliferate. Previous studies in our laboratory revealed that there are age-related differences in the proliferative capacity of HCECs.…”

mentioning

confidence: 99%

Age Differences in Cyclin-Dependent Kinase Inhibitor Expression and Rb Hyperphosphorylation in Human Corneal Endothelial Cells

Enomoto

Mimura

Harris

et al. 2006

Invest. Ophthalmol. Vis. Sci.

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p16(INK4a), p21WAF1/Cip1, p27Kip1, and Rb were all expressed in HCECs, regardless of donor age. Age-related differences in the relative expression of p16INK4a and p21WAF1/Cip1 and in the kinetics of Rb hyperphosphorylation led to the conclusion that, in addition to the normal inhibitory activity of p27Kip1, there is an age-dependent increase in negative regulation of the cell cycle by p16INK4a and p21WAF1/Cip1. This additional molecular mechanism may be responsible, at least in part, for the reduced proliferative response observed in HCECs from older donors.

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mentioning

confidence: 99%

Age Differences in Cyclin-Dependent Kinase Inhibitor Expression and Rb Hyperphosphorylation in Human Corneal Endothelial Cells

Enomoto

Mimura

Harris

et al. 2006

Invest. Ophthalmol. Vis. Sci.

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“…Although the HCECs have limited proliferative capacity in vivo , these cells have the ability to proliferate under in vitro culture conditions [ 18 , 25 , 29 , 30 , 31 , 32 , 36 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 ,…”

Section: Cell Sources For Tissue Engineering Of Corneal Endotheliumentioning

confidence: 99%

“…There are many different types of coating materials available for expansion of HCECs. These include collagen [ 28 , 56 , 65 , 74 , 90 , 93 , 97 , 99 , 110 , 139 ], fibronectin [ 59 , 90 , 93 , 97 , 110 ], gelatin [ 32 , 50 , 76 , 94 ], laminin [ 93 , 97 ], extracellular matrix (ECM) from cultured bovine corneal endothelial cells [ 25 , 27 , 59 , 67 , 71 , 93 ], a mixture of laminin and chondroitin sulfate [ 51 ], and a mixture of fibronectin, collagen, and albumin (FNC Coating Mix ® ) [ 97 , 101 ].…”

Section: Substrates For Cultivation Of Human Corneal Endothelial Cmentioning

confidence: 99%

Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium

Navaratnam

Utheim

Rajasekhar

et al. 2015

JFB

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Corneal endothelium is a single layer of specialized cells that lines the posterior surface of cornea and maintains corneal hydration and corneal transparency essential for vision. Currently, transplantation is the only therapeutic option for diseases affecting the corneal endothelium. Transplantation of corneal endothelium, called endothelial keratoplasty, is widely used for corneal endothelial diseases. However, corneal transplantation is limited by global donor shortage. Therefore, there is a need to overcome the deficiency of sufficient donor corneal tissue. New approaches are being explored to engineer corneal tissues such that sufficient amount of corneal endothelium becomes available to offset the present shortage of functional cornea. Although human corneal endothelial cells have limited proliferative capacity in vivo, several laboratories have been successful in in vitro expansion of human corneal endothelial cells. Here we provide a comprehensive analysis of different substrates employed for in vitro cultivation of human corneal endothelial cells. Advances and emerging challenges with ex vivo cultured corneal endothelial layer for the ultimate goal of therapeutic replacement of dysfunctional corneal endothelium in humans with functional corneal endothelium are also presented.

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“…Due to donor shortage, corneal endothelial tissue engineering is a promising alternative to corneal transplantation [ 3 ]. As early as 1990, Insler et al have reported a method to transplant corneal endothelial cells (CECs) expanded ex vivo on the carriers made of collagen-coated dextran [ 4 ]. In recent years, naturally occurring biopolymer such as chitosan has also been widely used to engineer CEC culture substrates.…”

Section: Introductionmentioning

confidence: 99%

Investigation of Overrun-Processed Porous Hyaluronic Acid Carriers in Corneal Endothelial Tissue Engineering

2015

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Hyaluronic acid (HA) is a linear polysaccharide naturally found in the eye and therefore is one of the most promising biomaterials for corneal endothelial regenerative medicine. This study reports, for the first time, the development of overrun-processed porous HA hydrogels for corneal endothelial cell (CEC) sheet transplantation and tissue engineering applications. The hydrogel carriers were characterized to examine their structures and functions. Evaluations of carbodiimide cross-linked air-dried and freeze-dried HA samples were conducted simultaneously for comparison. The results indicated that during the fabrication of freeze-dried HA discs, a technique of introducing gas bubbles in the aqueous biopolymer solutions can be used to enlarge pore structure and prevent dense surface skin formation. Among all the groups studied, the overrun-processed porous HA carriers show the greatest biological stability, the highest freezable water content and glucose permeability, and the minimized adverse effects on ionic pump function of rabbit CECs. After transfer and attachment of bioengineered CEC sheets to the overrun-processed HA hydrogel carriers, the therapeutic efficacy of cell/biopolymer constructs was tested using a rabbit model with corneal endothelial dysfunction. Clinical observations including slit-lamp biomicroscopy, specular microscopy, and corneal thickness measurements showed that the construct implants can regenerate corneal endothelium and restore corneal transparency at 4 weeks postoperatively. Our findings suggest that cell sheet transplantation using overrun-processed porous HA hydrogels offers a new way to reconstruct the posterior corneal surface and improve endothelial tissue function.

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Microcarrier cell culture of neonatal human corneal endothelium

Cited by 16 publications

References 11 publications

Age Differences in Cyclin-Dependent Kinase Inhibitor Expression and Rb Hyperphosphorylation in Human Corneal Endothelial Cells

Age Differences in Cyclin-Dependent Kinase Inhibitor Expression and Rb Hyperphosphorylation in Human Corneal Endothelial Cells

Substrates for Expansion of Corneal Endothelial Cells towards Bioengineering of Human Corneal Endothelium

Investigation of Overrun-Processed Porous Hyaluronic Acid Carriers in Corneal Endothelial Tissue Engineering

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