11. Anchor securely the hooks with cell construct onto the cham-

ber of the bioreactor by tightening the screws on two ends. Fill

up the chamber with complete medium. Cut the hooks con-

necter by sterile scissors. Put the glass lid on the chamber.

Culture overnight at 37
C with 5% CO2 (see Note 8).

12. Connect the wires for the actuator. Switch on the power and

corresponding channel controller to start mechanical stimula-

tion. Check the indicator lights and assure the bioreactor can

function properly. Move the bioreactor into the incubator and

subject the 3D cell construct to mechanical stretching for

6 days (stretch program: 6% stretching, 0.25 Hz, 8 h, followed

by 16 h rest).

3.2.2

Scaffold-Based

Mechanical Stimulation

1. Take a 30 mm  40 mm CelGro^® scaffold and make the rough

side of CelGro^® up. Put it on the bottom part of the culture

chamber. Firmly assemble the culture chamber (see Fig. 2b) (see

Note 9).

2. Thaw up the cells (mice TDSCs, one million cells, passage 2–4,

to 37
C) as descripted in Subheading 3.2.1, steps 1 and 2.

3. Seed cells (Quantity: 4  10⁵) into CelGro^® culture chamber.

Put the culture chamber in a Petri dish and culture 24 h at

37
C with 5% CO2 for cell attachment in CelGro^®.

4. Roll the 3D CelGro^® collagen-tenocyte construct up to

25  5 mm (see Fig. 2b).

5. Use the sterile clamp to anchor the collagen-tenocyte construct

onto the chamber of the bioreactor by tightening the screws on

two ends. Fill up the chamber with complete medium. Put the

glass lid on the chamber. Culture overnight at 37
C with 5%

CO2.

Fig. 2 Schematic diagram of (a) scaffold-free model and (b) scaffold-based model

140

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