Table of Contents - WOC 2012

FP-RET-TH 27 (6)
Imaging Retinal Pigment Epithelial Proliferation Secondary to
Photocoagulation in Vivo by Polarization Sensitive Optical
Coherence Tomography
Lammer Jan (1) , Bolz Matthias (1) , Pircher Michael (2) , Hitzenberger Christoph
(2) (1)
, Schmidt-Erfurth Ursula
1. Department of Ophthalmology, Med Univ Vienna
2. Center for Medical Physics and Biomedical Engineering, Med Univ Vienna
Purpose: To image the RPE after macular laser and to monitor the healing
response over time in vivo in patients with DME using polarization sensitive
optical coherence tomography (PS-OCT).
Methods: Thirteen eyes underwent grid photocoagulation for diabetic
maculopathy. Retinal healing processes were continuously followed over
the course of 3 months. A PS-OCT prototype was used allowing detection
and measurement of the RPE-changes based on their specific polarization
scrambling qualities.
Results: After one day after photocoagulation, RPE changes were rather
subtle. In tissue-sensitive PS-OCT imaging, polarization scrambling columns
were found at the level of the RPE by week 1. During follow up, different healing
responses were seen in the polarization scrambling RPE-layer, ranging from
hyper-proliferation to focal atrophy.
Conclusion: As a valuable new imaging tool, PS-OCT revealed specific
morphologic changes in the outer retinal layers undetectable with intensitybased
spectral domain OCT. The increase in polarization scrambling tissue
over the course of 3 months indicates a more intense healing reaction and
proliferation of RPE cells than previously characterized in rodent studies.
These findings support the hypothesis that the benefit of grid and focal macular
laser photocoagulation may be driven by an increase in RPE metabolic activity.
FP-RET-TH 27 (7)
Innovative Autologous Stem Cell Therapy in Retinal Diseases
Soriano Horacio (1) , Moviglia Gustavo
1. Univiversity Maimonides and Clinica Oftalmologica de Alta Complejidad
We have treated patients with Diabetic retinopathy at proliferative stage with T
cell vaccination. It is an immune modulator therapy used in clinic since
1991 that help to develop the effector [Cellular and Molecular Immunology.
2004, 1(5):321-327]. Patients with proven diagnosis of Diabetes Type 2 and
proliferative retinopathy lesions under went to a leukocyte apheresis to obtain
their Mononuclear cells. The cell suspension was cultured during 4 days in
DMEM + swine retina lysate. Activated CD4 cells against retina were negative
selected using Monoclonal antibodies attached to magnetic immunobeads
(Clinimac Kits, Miltenyi, Germany). Selected lymphocytes were irradiated
and administered intravenously 12 consecutive times in escalating doses,
once every 4 weeks. No serious adverse events were observed and the
clinical preliminary results seem to show that treated patients stop the disease
progression and partially revert their retinal damage. More time and treated
patients are needed to establish the efficacy and efficiency of the proposed
method to treat this condition.
WOC2012 Abstract Book
FP-RET-TH 27 (8)
Human Retinal Pigment Epithelial Cell Culture on polyHEMA Coated
Culture Substrates and the Pattern of Specific Gene Expression
Nazemroaya Fatemeh (1) , Soheili Zahra-Soheila (1) , Deezagi Abdolkhalegh (1) ,
Samiei Shahram (2) , Ahmadieh Hamid (3)
1. National Institute of Genetic Engineering and Biotechnology
2. Iranian Blood Transfusion Organization Research Center
3. Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences
Objective: Retinal pigment epithelium (RPE) is the pigmented cell layer
between photoreceptor cells and the choroiocapillares of the retina. poly-2hydroxyethyl
methacrylate (polyHEMA) is used to make soft contact lenses.
Cultured RPE cells on polyHEMA don›t adhere to substratum and join together
to form colonies.
Method: RPE cells were isolated from neonatal human globes and cultured in
DMEM/F12 containing 10% FBS. PolyHEMA (12 mg/ml) in 95% ethanol
was prepared and RPE cells were re-cultured on polyHEMA and polytstren
substrates (as control). 2 days later MTT assay (3-(4, 5-Dimethylthiazol-2yl)-2,5-diphenyltetrazolium
bromide) was applied to assess the number of
cells. RNA extraction and cDNA synthesis were performed to analyze gene
expression by Real-Time PCR.
Result: The number of RPE cells on polyHEMA and polystren were the same
and the expression of PAX6, CHX10-1, VSX-1, Tyrosinase, Thy1, RPE65,
CRABP1, CRABP2, Nestin GAPDH, PKC? genes were detected in cultured
RPE on polyHEMA.
Conclusion: Cells cultured on PolyHEMA can survive, form giant, pigmented
colonies and be re-cultured. The cultures expressed progenitor cell markers
and specific markers for terminally differentiated retinal cells. Probably this
polymer would be encouraging for maintenance and expansion of the retinal
progenitors/stem cells.
FP-RET-TH 27 (9)
Culture of Human Retinal Pigment Epithelial Cells in 3D-alginate
Polymer and Investigation of Viability and Gene Expression
Heidari Razeih (1) , Soheili Zahra-Soheila (1) , Deezagi Abdolkhalegh (1) , Ahmadieh
Hamid (2) , Samiei Shahram (3)
1. National Institute of Genetic Engineering and Biotechnology
2. Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences
3. Iranian Blood Transfusion Organization Research Center
Objective: RPE cells protect and nourish the retina, especially photoreceptor
cells. Alginate is extensively used in tissue engineering. It can form 3D-hydro
gel that living cells will be encapsulated in. This study evaluated the expression
of specific genes and viability of the cells in cultured RPE cells in alginate
hydro gel.
Method: RPE cells were isolated from neonatal human globes and cultured in
DMEM/F12 containing 10% FBS. Cells from passages 2-6 were cultured in
alginate beads. MTT (3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide)assay was applied to assess the number of cells . RNA extraction and
cDNA synthesis were done to analysis gene expression by Real-Time PCR
Result Viability of cells on alginate and polysterene were nearly the same.
Real-Time PCR data indicated cells in alginate express tyrosinase, RPE65,
thy1, nestin, PKC alpha, CRABP1-2, Pax6, CHX10 and VSX-1 genes.
Conclusion: The cells survived on alginate, could be re-cultured on polysteren
and performed a monolayer cells. In this condition, RPE cells retained
expression of their progenitor and specific genes. Our data indicated alginate
could be used as a scaffold for maintenance and expansion of RPE cells.
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