Acrobiosystems Co., Ltd. Class A · SZSE:301080
Acrobiosystems Co., Ltd. Class A — Technical Presentation (2023)
10 slides from Acrobiosystems Co., Ltd. Class A's technical presentation, 2023
Slide 1: Human Laminin 521 GMP Protein Stem Cell Culture Protocol
Human Laminin 521 GMP Protein, (Cat. No. GMP-LA5H24) Stem Cell Culture Protocol Scan this QR code for special gifts & more! GMP-LA5H24-P01-Uni-V01
Slide 2: 1. Product Background
1. Product Background Laminin 521 (LN521) is a recombinant human protein that provides a defined surface for in vitro feeder-free culture of multiple human pluripotent stem cells (PSCs). LN521 has been proven to maintain normal growth characteristics and stemness in multiple PSC lines without simult
Slide 3: 2. Experimental instruments and materials
2. Experimental instruments and materials 2.1 Instruments: Biosafety cabinet, cell incubator, low temperature horizontal centrifuge, inverted microscope. 2.2 Materials: Sterile pipette tips; Sterile EP tube and other consumables. 2.3 Reagents: Laminin-521(GMP-LA5H24), ROCK inhibitor(Y-27632), DMEM F
Slide 4: 3. Experimental contents and methods
3. Experimental contents and methods 3.1. Laminin plate coating: 3.1.1. Use sterile deionized water or water for injection to reconstitute laminin-521 to 400µg/ml. It is recommended to further dilute to 100µg/ml with sterile D-PBS(Ca++/Mg++) prior to use, and then dilute to working concentration of
Slide 5: Culture vessel Coating
Culture Coating Volume Volume Total mTeSR- vessel concentration （laminin） （DPBS） volume plus media 6-well 0.5 µg/cm2 50 μL/well 950 μL/well 1 mL/well 2 mL 12-well 0.5 µg/cm2 25 μL/well 475 μL/well 0.5 mL/well 1 mL 24-well 0.5 µg/cm2 15 μL/well 285 μL/well 0.3 mL/well 0.5 mL 35-mm 0.5 µg/cm2 50 μL/we
Slide 6: 4. Cell Culture - iPSC
3.1.2. Add indicated volumes of the laminin-DPBS mixture into the each well and gently shake to ensure that matrix is spread across the well. 3.1.3. Transfer the plate into a 4°C refrigerator for incubation overnight or 37°C incubator for at least 2 hours. Overnight is recommended for ideal cell cul
Slide 7: 4.1.5 Cell plating and iPSC passaging
4.1.5. Discard the well-coated laminin solution in 12 well plate. 4.1.6. Discard the cell supernatant and gently resuspend iPSCs with 1 mL mTeSR-plus containing 10 µM Y-27632, and transfer it to a coated 12 well plate. Shake plate to evenly distribute cells, and stand at room temperature for 10 minu
Slide 8: 4.2.2 iPSC passaging protocol
4.2.2. Discard the PBS and add 0.5 mL ReLeSR. Stand at 37℃ for 8-10 minutes or keep under microscopic observation until cells are not bound to the plate. NOTE: For the passage of induced pluripotent stem cells (iPSCs), TrypLE Select, ReLeSR, and Accutase are also recommended. When dissociating iPSCs
Slide 9: 4.2.6 Cryopreservation of iPSCs
4.2.6. Transfer the cells to the prepared laminin-coated 12-well plate. Shake the plate gently to evenly distribute cells, and stand at room temperature for 10-20 minutes. 4.2.7. Place the 12-well plate into a 37°C incubator for incubation. 4.3. Cryopreservation of iPSCs 4.3.1. Prepare ReLeSR for us
Slide 10: Scan this QR code for special gifts & more!
Scan this QR code for special gifts & more! Copyright © 2023 ACROBiosystems All rights reserved.