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Table 3 Generation of immature RAPA-DC

From: Immunoregulatory properties of rapamycin-conditioned monocyte-derived dendritic cells and their role in transplantation

 

Research

Clinical

Monocyte isolation

CD14 microbeads (AutoMACS® or CliniMACS®)

Elutriation or CD14 microbeads (CliniMACS®)

Culture conditions

10% fetal calf serum

10% human serum

(7 days)

AIM-V media

AIM-V media (c-GMP grade)

 

1000 U/mL rhIL-4

1000 U/mL rhIL-4 (c-GMP grade)

 

1000 U/mL GM-CSF

1000 U/mL GM-CSF (c-GMP grade)

 

10 ng/mL Rapamycin

10 ng/mL Rapamycin (c-GMP grade)

 

Cell culture plates

Cell culture plates or Aastrom Replicell system