All-inclusive 3D Human Retinal Pigment Epithelial Spheroid Formation Kit

Age-related macular degeneration (AMD) is characterized in its early stages by the presence of extracellular deposits, known as drusen, that accumulate between the basal surface of the retinal pigmented epithelium and Bruch's membrane, an extracellular matrix complex that separates the neural retina from the capillary network in the choroid [1]. Several studies have shown that drusen contains a variety of protein and lipid components [2]. Although liver is the primary biosynthetic site for most of these molecules, retinal pigment epithelial (RPE) cells locally synthesize a number of drusen components [2]. The respective contributions of RPE-derived and plasma-derived molecules to the biogenesis of drusen, and the relevant molecular interactions leading to drusen depositions, however, have not been fully identified. One of the major limitations is that RPE cells, once isolated from the eye, tend to dedifferentiate into myofibroblasts in conventional 2D cell culture. Recent studies have shown that 3D retinal pigment epithelial cell spheroids form and maintain a well-differentiated epithelium in 3D cell culture [3]. ScienCell develops an all-inclusive 3D human retinal pigment epithelial spheroid formation kit (3D-HRPEpiSF). 3D RPE spheroids exhibit the differentiated epithelial cell marker cytokeratin-18 and deposit apolipoprotein ApoE, a prominent drusen constituent. The 3D RPE spheroid model is an ideal way to model drusen in vitro and study the pathogenesis of related diseases, such as AMD.

Kit Components:

3D Cell Culture Components

• 1 x #6540 - Human Retinal Pigment Epithelial Cells (HRPEpiC) - 5 x; 10^5 cells - Storage: Liquid nitrogen
• 1 x #3D-4101 - 3D-Epithelial Spheroid Medium (3D-EpiSpM) - 200 mL - Storage: 2-8°C
• 1 x #3D-4152 - 3D-Epithelial Spheroid Supplement (3D-EpiSpS) - 2 mL - Storage: -20°C
• 1 x #0004 - Fetal Bovine Serum (FBS) - 4 mL - Storage: -20°C
• 1 x #0583 - Penicillin/Streptomycin Solution (P/S) - 2 mL - Storage: -20°C
• 1 x #0343 or #0353 or #0383 - Ultra-Low Binding Culture Plates (24-, 48-, or 96- well plate) - 1 plate - Storage: RT

2D Cell Culture Components

• 1 x #4101 - Epithelial Cell Medium (EpiCM) - 500 mL - Storage: 2-8°C
• 1 x #4152 - Epithelial Cell Growth Supplement (EpiCGS) - 5 mL - Storage: -20°C
• 1 x #0010 - Fetal Bovine Serum (FBS) - 10 mL - Storage: -20°C
• 1 x #0503 - Penicillin/streptomycin Solution (P/S) - 5 mL - Storage: -20°C

Additional Recommended Materials (Not Included):

• #0113 - Trypsin Neutralization Solution
• #0183 - 0.05% Trypsin/EDTA (T/E)
• #0303 - Dulbecco's Phosphate-Buffered Saline (DPBS)
• #0413 - Poly-L-Lysine (10 mg/mL)

ScienCell's 3D human retinal pigment epithelial spheroids display the differentiated epithelial cell marker cytokeratin-18 and deposit apolipoprotein ApoE, a prominent drusen constituent. The 3D RPE spheroid model is an ideal way to model drusen in vitro and study the pathogenesis of related diseases, such as AMD.