Rpmi based media

Bone marrow-derived
dendritic cells (BMDCs) were derived using standard techniques. In
brief, bone marrow cells were isolated from female C57Bl/6J mice (Jackson
Laboratories) and cultured in RPMI based media (Lonza) supplemented
with 10% heat inactivated FBS (Sigma-Aldrich), 1% penicillin/streptomycin,
50 μM β-mercaptoethanol, and 20 ng/mL GM-CSF (Peprotech).
BMDCs were harvested and used for experiments between day 7 and 10
of differentiation. Differentiation was confirmed using the CD11c,
CD11b and MHC-II surface markers. For the in vitro cell infiltration study, 2 × 10⁶ BMDCs dispersed
in 50 μL of cell culture media were seeded on top of the scaffold
(PLG-9, PLG-14, and PLG-18) and the scaffold was incubated at 37 °C
for 1 h to allow cell infiltration into the macropores of the scaffold.
Then, the surface of the scaffolds was carefully rinsed with a culture
media to remove the uninfiltrated cells. To retrieve the infiltrated
cells from the scaffold, the scaffold was cut with a sterile razor
blade into small pieces and subsequently digested with collagenase
type II (Worthington, 250 U/mL) for 1 h at 37 °C to detach any
attached cells to the scaffold matrix.^{11 (link),18 ,20 (link)} The cells were collected by filtration of the mixture
of cells and PLG pieces using 40 μm cell strainers. The retrieved
cells were counted using a Coulter counter.

BMDC were derived using standard techniques^{49 (link),50 (link)}. In brief, bone marrow cells were isolated from female C57Bl/6J mice (Jackson Laboratories) and cultured in RPMI based media (Lonza) supplemented with 10% heat inactivated FBS (Sigma-Aldrich), 1% penicillin/streptomycin, 50µM β-mercaptoethanol and 20 ng/ml GM-CSF (Peprotech). Dendritic cells were harvested and used for experiments between day 7 and 10 of differentiation. Differentiation was confirmed using the CD11c, CD11b and MHC II surface markers.