Purpose: This study focuses on the development of the method of introduction of magnetic microparticles in the cytoplasm of HEK-293 cell line with their subsequent fixation under the retina of the eye.Materials and Methods. Magnetic particles (d = 2,8 mm) were treated with pluronic and injected into the cytoplasm of HEK-293 cell line, expressing GFP. The surgery was made under general anesthesia. HEK-293 containing magnetic particles were injected into the subretinal space of rabbit eyes (eyes 96, 48 rabbits) using original dosing device. In the experimental group (48 eyes, 24 rabbits) we fixed episcleral magnetic implant to hold cells in local place. In the control group (48 eyes, 24 rabbit) magnetic implant was not fixed. After the surgery all animals were examined using biomicroscopy, ophthalmoscopy with photographic recording, ultrasound, computed tomography and morphological study in certain terms (1, 3, 5, 7, 14, 21 day and 1 month).Results: The introduction of the magnetic particles into the cytoplasm of HEK 293 cell line has no effect on cell viability. HEK-293 containing magnetic particles remains in the place of injection during 21 days in rabbit eyes, where the magnetic implants were fixed (in control group during 3 days). Conclusions: Using of cells containing magnetic particles with fixation of the magnetic implant can be a promising method for cell therapy for the treatment of retinal diseases.
Purpose. To develop a technique for local subretinal injection of xenogeneic stem cells labeled with magnetic particles and to prove experimentally its effectiveness.Material and methods. We used a line of stem cells HEK-293 GFP, labeled with magnetic particles. The study was made in 84 eyes of 42 chinchilla rabbits aged 6 months, the weight was from 2.5 to 3.5kg. All right eyes were experimental (42 eyes) and all left eyes (42 eyes) were in the control group. In the experimental group we used original complex of polymer elastic magnetic implant (PEMI) with a laser probe and fixed it to the sclera, then we made a median vitrectomy and injected HEK-293 GFP under the retina using a specially designed dispenser. In the control group PEMI was not fixed. We examined animals using biomicroscopy, ophthalmoscopy, ultrasound scanning, optical coherence tomography (OCT), computer tomography (CT), morphological study (cryo-histological sections) 1, 3, 5, 7, 14 days and 1 month after surgery.Results. According to the results of biomicroscopy in observation periods up to 3 days the vascular injection was visualized in the area operation. According to the results of ophthalmoscopy and ultrasound scanning during the first day the local retinal detachment was visualized in the area of local injection of the stem cells, which was not noted in terms of further observations. CT helped us to confirm the localization of PEMI fixation. The morphological study results showed that cells were located in the subretinal space up to 14 days in the experimental group, and only up 3 days in the control group.Conclusion. The developed surgical technique enables to control the injection of cells into the subretinal space, reduces the risk of tissue damage and exit cells in the vitreous space. The suggested method allows to fix the cellular material in the local place of the injection and enables to predict cells` movement.
Purpose: is to develop a technique for local subretinal injection of xenogeneic stem cells labeled with magnetic particles and to prove experimentally its effectiveness.Material and methods: We used a line of stem cells HEK-293 GFP,labeled with magnetic particles. The study was made on 84 eyes of 42 chinchilla rabbits 6 months of age, the weight were from 2.5 to 3.5 kg. All right eyes were experimental (42 eyes) and all left eyes (42 eyes) were the control group. In the experimental group we used original complex of polymer elastic magnetic implant (PEMI) with laser probe and fixed it to the sclera, then we made a median vitrectomy and injected HEK-293 GFP under the retina using a specially designed dispenser. In the control group PEMI was not fixed. We examined animals using biomicroscopy, ophthalmoscopy, ultrasound scanning, optical coherence tomography OCT), computer tomography (CT), morphological study (cryohistological sections) in 1, 3, 5, 7, 14 day and 1 month after surgery.Results: According the results of biomicroscopy in observation periods up to 3 days the vascular injection was visualized in the area operation. According the results of ophthalmoscopy and ultrasound scanning in 1 day the local retinal detachment was visualized in the area of local injection of the stem cells, which was not visualized in terms of further observations. CT helped us to confirm the local place of PEMI fixation. The morphological study results showed that cells were located in the subretinal space up to 14 days in the experimental group, and only up 3 days in the control group.Conclusion: The suggested surgical technique enables to control the injection of cells into the subretinal space, reduces the risk of tissue damage and exit cells in the vitreous space. The suggested methodology allows the fixing of the cellular material in the local place of the injection and enables to predict cells`s movement.
Purpose : is to develop a technique for local subretinal injection of xenogeneic stem cells labeled with magnetic particles and to prove experimentally its effectiveness. Material and methods: We used a line of stem cells HEK-293 GFP, labeled with magnetic particles. The study was made on 84 eyes of 42 chinchilla rabbits 6 months of age, the weight were from 2.5 to 3.5 kg. All right eyes were experimental (42 eyes) and all left eyes (42 eyes) were the control group. In the experimental group we used original complex of polymer elastic magnetic implant (PEMI) with laser probe and fixed it to the sclera, then we made a median vitrectomy and injected HEK-293 GFP under the retina using a specially designed dispenser. In the control group PEMI was not fixed. We examined animals using biomicroscopy, ophthalmoscopy, ultrasound scanning, optical coherence tomography OCT), computer tomography (CT), morphological study (cryohistological sections) in 1, 3, 5, 7, 14 day and 1 month after surgery. Results: According the results of biomicroscopy in observation periods up to 3 days the vascular injection was visualized in the area operation. According the results of ophthalmoscopy and ultrasound scanning in 1 day the local retinal detachment was visualized in the area of local injection of the stem cells, which was not visualized in terms of further observations. CT helped us to confirm the local place of PEMI fixation. The morphological study results showed that cells were located in the subretinal space up to 14 days in the experimental group, and only up 3 days in the control group. Conclusion: The suggested surgical technique enables to control the injection of cells into the subretinal space, reduces the risk of tissue damage and exit cells in the vitreous space. The suggested methodology allows the fixing of the cellular material in the local place of the injection and enables to predict cells`s movement.
Purpose: This study focuses on the development of the method of introduction of magnetic microparticles in the cytoplasm of HEK-293 cell line with their subsequent fixation under the retina of the eye. Materials and Methods. Magnetic particles (d = 2,8 mm) were treated with pluronic and injected into the cytoplasm of HEK-293 cell line, expressing GFP. The surgery was made under general anesthesia. HEK-293 containing magnetic particles were injected into the subretinal space of rabbit eyes (eyes 96, 48 rabbits) using original dosing device. In the experimental group (48 eyes, 24 rabbits) we fixed episcleral magnetic implant to hold cells in local place. In the control group (48 eyes, 24 rabbit) magnetic implant was not fixed. After the surgery all animals were examined using biomicroscopy, ophthalmoscopy with photographic recording, ultrasound, computed tomography and morphological study in certain terms (1, 3, 5, 7, 14, 21 day and 1 month). Results: The introduction of the magnetic particles into the cytoplasm of HEK 293 cell line has no effect on cell viability. HEK-293 containing magnetic particles remains in the place of injection during 21 days in rabbit eyes, where the magnetic implants were fixed (in control group during 3 days). Conclusions: Using of cells containing magnetic particles with fixation of the magnetic implant can be a promising method for cell therapy for the treatment of retinal diseases.