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. 2022 Apr 24;9(18):2104451. doi: 10.1002/advs.202104451

Table 3.

Overview of different types of enzymatic digestion solutions and cell culture media used in this study

Name of solution/medium Use Ingredients
Adipose tissue digestion solution
  • –

    Digestion of adipose tissue for adipocyte and SVF isolation

 DMEM/F‐12 + 0.13 U mL−1 collagenase type NB4 + 1% (w/v) BSA
A wash medium
  • –

    Wash adipocytes during isolation

  • –

    Wash adipocytes prior to injection

 DMEM/F‐12 + 100 U mL−1 Pen/Strep
A base medium
  • –

    Overnight flask culture of adipocytes (A) between isolation and chip injection

 DMEM/F‐12 (incl. glutamine) + 10% (v/v) FCS + 10 µM HEPES + 100 U mL−1 Pen/Strep
S/M base medium
  • –

    Overnight flask culture of SVF/CD14+ cells between isolation and chip injection

  • –

    First seeding of PBMC‐derived CD14+‐cells

 A base medium + 10 ng mL−1 GM‐CSF
Skin digestion solution
  • –

    Digestion of dissected skin for mvEC isolation

 PBS−+ 2.0 /mL Dispase II
mvEC expansion medium
  • –

    Expansion of isolated mvECs in cell culture flask format

 ECGM + 10 mg mL−1 Gentamicin
PBMC culture medium
  • –

    For PBMC thawing process

 X‐VIVO 15 + 10% (v/v) autologous serum + 100 U mL−1 Pen/Strep
PBMC freezing medium
  • –

    For freezing PBMCs after isolation

 10% (v/v) dimethylsulfoxid (DMSO) in FCS
PBMC thawing medium
  • –

    Overnight plate culture of thawed PBMCs

 PBMC culture medium + 10 U mL−1 IL‐2
T cell activation medium
  • –

    In plate culture medium for activation of thawed PBMCs prior to chip loading

 PBMC culture medium + 1% (v/v) T Cell TransAct human
2× Monocyte/macrophage culture medium
  • –

    Maintenance of PBMC‐derived CD14+ cells for 5 days

 A base medium + 20 ng mL−1 GM‐CSF
A medium
  • –

    On‐chip cultures of adipocyte (A)‐only chips

 A base medium + 60 nM Insulin + 100 nM Rosiglitazone
E medium
  • –

    On‐chip cultures of mvEC (E) layer‐only chips

 ECGM + 100 U mL−1 Pen/Strep
E TNF‐α stimulation medium
  • –

    24 h pro‐inflammatory stimulation with TNF‐α of mvEC (E) layer‐only chips

 E medium + 20 ng mL−1 TNF‐α
E LPS stimulation medium
  • –

    24 h pro‐inflammatory stimulation with LPS of mvEC (E) layer‐only chips

 E medium + 100 ng mL−1 LPS
AE co‐culture medium
  • –

    On‐chip co‐cultures of adipocytes (A) and endothelial layer (E)

 A medium:E medium 1:1
AS/AM co‐culture medium
  • –

    On‐chip co‐cultures of adipocytes (A) and SVF (S) or CD14+‐cells (M)

  • –

    Perfusion of monocytes

 A medium + 10 ng mL−1 GM‐CSF
ASE multi‐culture medium
  • –

    On‐chip multi‐culture of adipocytes (A), endothelial layer (E), and SVF (S)

 AE co‐culture medium + 10 ng mL−1 GM‐CSF
A TNF‐α stimulation medium
  • –

    24 h pro‐inflammatory stimulation with TNF‐α of adipocyte (A)‐only chips

 A base medium + 20 ng mL−1 TNF‐α
AS/AM TNF‐α stimulation medium
  • –

    24 h pro‐inflammatory stimulation with TNF‐α of on‐chip co‐cultures of adipocytes (A) and SVF (S) or CD14+‐cells (M)

 A base medium + 10 ng mL−1 GM‐CSF + 20 ng mL−1 TNF‐α
ASE TNF‐α stimulation medium
  • –

    24 h pro‐inflammatory stimulation with TNF‐α of on‐chip multi‐culture of adipocytes (A), endothelial layer (E), and SVF (S)

 A base medium:E medium 1:1 + 10 ng mL−1 GM‐CSF + 20 ng mL−1 TNF‐α
A LPS stimulation medium
  • –

    24 h pro‐inflammatory stimulation with LPS of adipocyte (A)‐only chips

 A base medium + 100 ng mL−1 LPS
AS/AM LPS stimulation medium
  • –

    24 h pro‐inflammatory stimulation with LPS of on‐chip co‐cultures of adipocytes (A) and SVF (S) or CD14+‐cells (M)

 A base medium + 10 ng mL−1 GM‐CSF + 100 ng mL−1 LPS
ASE LPS stimulation medium
  • –

    24 h pro‐inflammatory stimulation with LPS of on‐chip multi‐culture of adipocytes (A), endothelial layer (E), and SVF (S)

 A base medium:E medium 1:1 + 10 ng mL−1 GM‐CSF + 100 ng mL−1 LPS
T‐cell perfusion medium
  • –

    Overnight perfusion of circulating immune cells

 X‐VIVO 15 + 100 U mL−1 Pen/Strep