Table 2 ECM coating conditions used in this study.

From: Polyethyleneimine facilitates the growth and electrophysiological characterization of iPSC-derived motor neurons

Abbreviation

ECM components

References

Cost(€)/ml*

POL

20 μg/ml Poly-L-ornithine + 20 μg/ml Laminin

9

5.15

POLF

20 μg/ml Poly-L-ornithine + 20 μg/ml Laminin

 + 10 μg/ml Fibronectin

10

8.23

POLFM

20 μg/ml Poly-L-ornithine + 20 μg/ml Laminin + 10 μg/ml Fibronectin + Matrigel (1:20)

12

11.38

POM

20 μg/ml Poly-L-ornithine + Matrigel (1:50)

11

1.79

PEI

0.1% Polyethyleneimine in borate buffer

 

0.11

  1. * The detailed calculation is listed in Table S1.
  2. POL: 100 µg/ml Poly-L-ornithine (P4957, Sigma) was diluted to 20 µg/ml in Dulbecco’s Phosphate Buffered Saline (DPBS, D8662, Sigma). The culture ware was coated with 20 µg/ml Poly-L-ornithine at 4℃ overnight. Laminin at 1 mg/ml (L2020, Sigma) was thawed at 4℃ overnight and diluted to 20 µg/ml in DPBS. On the second day of coating, Poly-L-ornithine was removed, and the plates were rinsed with sterile water twice and then coated with 20 µg/ml of Laminin at 37 °C for 2 h. The coating matrix was removed and rinsed once with PBS before plating cells.
  3. POLF: The dishes were first coasted with poly-l-ornithine as described above. The 1 mg/ml laminin and 1 mg/ml fibronectin (F1141, Sigma) stocks were pre-thawed at 4℃ and diluted with DPBS to contain 20 μg/ml laminin and 10 μg/ml fibronectin. Then poly-l-ornithine pre-coated plates were coated with laminin/fibronectin solution at 37 °C for 2 h. The coating matrix was removed and rinsed once with PBS before plating cells.
  4. POLFM: After poly-l-ornithine, laminin and fibronectin coating as described above, Matrigel was thawed at 4℃ and diluted 20 × in KnockOut™ DMEM (10,829,018, Gibco), and then added to the same wells. The plates were kept at 37 °C for 2 h, and Matrigel was removed before use.
  5. POM: After overnight coating of poly-l-ornithine, the same wells were coated with Matrigel (1:50) at 37 °C for 2 h, and Matrigel was removed before use.
  6. 0.1% PEI Solution: 50% PEI (P3143, Sigma) was diluted to 10% with sterile water as recommended by FujiFilm Cellular Dynamics, Inc. (https://www.fujifilmcdi.com/icell-motor-neurons-01279-gmnc01279). The 0.1% PEI working solution was freshly made by diluting 10% PEI at a 1: 100 ratio with 1 × borate buffer, which was made by diluting 20 × borate buffer (28,341, Thermo Scientific) with sterile water. The 0.1% PEI was filtered with a 0.22 μM sterile filter (A16534K, Lennox) before use.
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