研究目的
Investigating the survival and integration of expanded pig retinal progenitor cells (pRPCs) in normal recipients with and without transient anti-inflammatory suppression.
研究成果
Local transient rapamycin/dexamethasone immunosuppressive treatment had no significant impact on allogeneic survival and engraftment of pRPCs transplanted into normal wild-type pigs with healthy retina. Grafted cells maintained the ability to differentiate into photoreceptors after 4 weeks in vivo.
研究不足
The study did not investigate longer time points or transplantation into hosts with retinal degeneration.
1:Experimental Design and Method Selection:
pRPCs were derived from the neural retina of E60 GFP transgenic pigs, expanded for six passages, characterized, and transplanted into the subretinal space of 12 pigs. Six recipients received a single intravitreal injection of rapamycin and dexamethasone.
2:Sample Selection and Data Sources:
pRPCs were isolated from E60 embryos of EGF-transgenic pigs.
3:List of Experimental Equipment and Materials:
Collagenase IV, fibronectin-coated flasks, Ultraculture medium, rh bFGF, rh EGF, Trypzean, DTI, benzonase, HBSS-NAC, rapamycin, dexamethasone.
4:Experimental Procedures and Operational Workflow:
Cells were passaged upon reaching 80% confluency, formulated as a suspension of 50,000 cells/μL in HBSS-NAC, and transplanted into the subretinal space.
5:Data Analysis Methods:
Immunocytochemistry, RT-PCR, histology, and fluorescence immunohistochemistry were used to analyze the results.
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Collagenase IV
Sigma-Aldrich
Digestion of neuroretinas
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fibronectin-coated flasks
Akron
Culture of pRPCs
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Ultraculture medium
Culture medium for pRPCs
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rh bFGF
Peprotech
Supplement for culture medium
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rh EGF
Supplement for culture medium
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Trypzean
Sigma-Aldrich
Passaging of pRPCs
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DTI
Gibco
Passaging of pRPCs
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benzonase
EMD Chemicals
Passaging of pRPCs
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HBSS-NAC
Formulation of pRPC suspension
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rapamycin
Immunosuppressive treatment
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dexamethasone
Immunosuppressive treatment
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