2.8. Cell-Laden Hydrogel Construct Subcutaneous Implantation in Nu/Nu Mice

Constructs with tethered TGF-β3 were thrice rinsed in PBS prior to implantation in athymic (nu/nu) mice (Charles River). Mice were anesthetized with isofluorane. Two incisions were made to form dorsal subcutaneous pockets above each limb. A total of four pockets were prepared each with a single implant (4 implants/mouse). Incisions were closed with staples that were removed after 10 days. After 15, 43, and 64 days mice were euthanized via CO₂ asphyxiation and cervical dislocation. All protocols followed were approved by the CU Boulder Institutional Animal Care and Use Committee and follow all guidelines from the NIH.