IN VITRO DIFFERENTIATION OF HUMAN BONE MARROW STEM CELLS INTO RETINAL PIGMENT EPITHELIUM
Hani Mohammed MohammedShaheen;
Abstract
SUMMARY
B
ackground and objective: retinal pigment epithelium (RPE) is a monolayer of cells between the neural retina and the choriocapillaris, plays crucial roles in the maintenance and function of the retina and its photoreceptors. These include the formation of the blood retinal barrier, absorption of stray light, supply of nutrients to the neural retina, regeneration of visual pigment, and uptake and recycling of shed outer segments of photo-receptors.
Dysfunction, degeneration, and loss of RPE cells are prominent features of subtypes of retinitis pigmentosa (RP) which is a cause of visual disability. In this condition, there is progressive visual loss that often leads to blindness. A variety of therapeutic approaches to delay the degenerative process are under development, but the grim reality is that many patients eventually lose their sight.
So, in the present studybone marrow stem cells was examined for the possibility of its differentiation into RPE in vitro.
Aim: this study was aimed to examine whether bone marrow derived stem cells can be differentiatedinto retinal pigmented epithelial cells in vitro after specific treatment or notand this might give the hope for treatment and regeneration of degenerated retinal cells as in cases of retinitis pigmentosa.
Material and method:Bone marrow was aspirated under local anesthesia from the posterior superior iliac spine. A total of 3 ml bone marrow aspirated on preservative free heparin from 20 patients was collected. Mononuclear fraction was collected using density gradient centrifugation. Mononuclear cells were plated in tissue culture flasks to multiply cells. Then Mesenchymal stem cell (MSC) characterization was done by flow cytometer for detecting CD90& CD271 on the cell surface and exclusion of CD34 which is hematopoietic cell marker. Then BMSCs were differentiated into RPE cells by adding Activin A and Nicotinamide to the culture. After thatRT-PCR was done to prove differentiation of cells into RPE cells.
Results:Bone marrow stem cells in all samples were cultured for 8 weeks then RT-PCR was done for the differentiated cells to detect mRNA gene responsible for expression of MiTF-A which is an early RPE marker. The results show that all samples were differentiated into RPE cells after 8 weeks of culture with activin A and nicotinamide.
B
ackground and objective: retinal pigment epithelium (RPE) is a monolayer of cells between the neural retina and the choriocapillaris, plays crucial roles in the maintenance and function of the retina and its photoreceptors. These include the formation of the blood retinal barrier, absorption of stray light, supply of nutrients to the neural retina, regeneration of visual pigment, and uptake and recycling of shed outer segments of photo-receptors.
Dysfunction, degeneration, and loss of RPE cells are prominent features of subtypes of retinitis pigmentosa (RP) which is a cause of visual disability. In this condition, there is progressive visual loss that often leads to blindness. A variety of therapeutic approaches to delay the degenerative process are under development, but the grim reality is that many patients eventually lose their sight.
So, in the present studybone marrow stem cells was examined for the possibility of its differentiation into RPE in vitro.
Aim: this study was aimed to examine whether bone marrow derived stem cells can be differentiatedinto retinal pigmented epithelial cells in vitro after specific treatment or notand this might give the hope for treatment and regeneration of degenerated retinal cells as in cases of retinitis pigmentosa.
Material and method:Bone marrow was aspirated under local anesthesia from the posterior superior iliac spine. A total of 3 ml bone marrow aspirated on preservative free heparin from 20 patients was collected. Mononuclear fraction was collected using density gradient centrifugation. Mononuclear cells were plated in tissue culture flasks to multiply cells. Then Mesenchymal stem cell (MSC) characterization was done by flow cytometer for detecting CD90& CD271 on the cell surface and exclusion of CD34 which is hematopoietic cell marker. Then BMSCs were differentiated into RPE cells by adding Activin A and Nicotinamide to the culture. After thatRT-PCR was done to prove differentiation of cells into RPE cells.
Results:Bone marrow stem cells in all samples were cultured for 8 weeks then RT-PCR was done for the differentiated cells to detect mRNA gene responsible for expression of MiTF-A which is an early RPE marker. The results show that all samples were differentiated into RPE cells after 8 weeks of culture with activin A and nicotinamide.
Other data
| Title | IN VITRO DIFFERENTIATION OF HUMAN BONE MARROW STEM CELLS INTO RETINAL PIGMENT EPITHELIUM | Other Titles | تحول الخلايا الجذعية المشتقة من نخاع العظام الى نسيج صباغى شبكى خارج الجسم | Authors | Hani Mohammed MohammedShaheen | Issue Date | 2014 |
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