Menicon ShellPa Pro, Instruction Manual, Page: 31 / 32

SP_Inst02 /02_20220208 Instruction Manual For ShellPa Pro (Model type : STMP) Page(31/32)
Lifescience Dept. Menicon Co.,Ltd.
Appendix 1
1.
Chamber coating procedure
(1) Coating with fibronectin
1) Dilute fibronectin with PBS to the concentration of 2 to 20 μg/mL.
2) Set a stretch chamber SC4Dea into a dish, add 1 to 1.5 mL of the diluted fibronectin solution 1) to
the chamber. Gently tap the dish to completely cover the bottom of the chamber.
3) Incubate the dishes for 30 minutes to 4 hours at 37°C.
(If the coating is not sufficient, please extent the incubation time.)
4) Remove the coating solution and wash 2-3 times with PBS or serum free culture medium. Seed the
cells to start culture.
(2) Coating with collagen
1) Prepare dilute hydrochloric acid (pH=3.1mM) and autoclave.
2) Dilute collagen (type I or type 4) with dilute hydrochloric acid 1).
3)
Set a stretch chamber SC4Dea into a dish, add 1 to 1.5 mL of the diluted collagen solution 2) to the
chamber. Gently tap the dish to completely cover the bottom of the chamber.
4) Incubate the dishes for 30 minutes to 4 hours at 37°C.
(If the coating is not sufficient, please extent the incubation time.)
5) Remove the coating solution and wash 2-3 times with PBS or serum free culture medium. Seed the
cells to start culture.
(3) Coating with laminin
1) Dilute laminin with PBS to the concentration of 1 μg/mL.
(Natural Mouse Laminin (Gibco #23017-015)
http://www.lifetechnologies.com/order/catalog/product/23017015 )
2) Set a stretch chamber SC4Dea into a dish, add 1 to 1.5 mL of the diluted laminin solution 1) to the
chamber. Gently tap the dish to completely cover the bottom of the chamber.
3) Incubate the dishes for 30 minutes to 4 hours at 37°C.
(If the coating is not sufficient, please extent the incubation time.)
4) Remove the coating solution and wash 2-3 times with PBS or serum free culture medium. Seed the
cells to start culture.
(4) Coating with PDL
1) Dilute PDL with serum free medium to the concentration of 4 mg/mL for storage at -20°C.
(Poly-D-Lysine Hydrobromide, High Molecular Weight (BD #354210))
2) Prior to use, dilute the stock PDL solution to the final concentration of 50 μg/mL.