Bronectin

The isolation procedure of endometrial cells was carried out as described [5] . The tissues were minced and digested with PBS containing collagenase type III (0.3 mg/ml, Worthington Biochemical Corporation, NJ, USA) and deoxyribonuclease type I (40 μg/ml, Worthington Biochemical Corporation) for one hour at 37 o C. Red blood cells were removed using Ficoll-Paque (GE Healthcare, Uppsala, Sweden) densitygradient centrifugation. Leukocytes were excluded using anti-CD45 antibody-coated Dynabeads (Invitrogen, Waltham, MA, USA). Epithelial cells were separated from the stromal cells using anti-CD326 (EpCAM) antibody-coated microbeads (Miltenyi Biotec Inc., San Diego, CA, USA). Freshly isolated epithelial cells and stromal cells were used for coculture and collection of condition medium as describe below. Some of the spare puri ed stromal cells (6000-8000 cells/cm 2 ) were plated into 100-mm petri dishes coated with bronectin (1 mg/ml, Invitrogen) and cultured in growth medium containing 10% FBS (Invitrogen), 1% antibiotics (Invitrogen) and 2 mmol/L glutamine (Invitrogen) in DMEM/F12 (Sigma-Aldrich, St Louis, MA, USA) for 7-14 days in a humidi ed carbon dioxide incubator at 37 o C in 5% CO 2 . The medium was changed every seven days until the cells reached 90% con uence.

The isolation procedure of endometrial cells was carried out as described (5) . The tissues were minced and digested with PBS containing collagenase type III (0.3 mg/ml, Worthington Biochemical Corporation, NJ, USA) and deoxyribonuclease type I (40 μg/ml, Worthington Biochemical Corporation) for one hour at 37 o C. Red blood cells were removed using Ficoll-Paque (GE Healthcare, Uppsala, Sweden) densitygradient centrifugation. Leukocytes were excluded using anti-CD45 antibody-coated Dynabeads (Invitrogen, Waltham, MA, USA). Epithelial cells were separated from the stromal cells using anti-CD326 (EpCAM) antibody-coated microbeads (Miltenyi Biotec Inc., San Diego, CA, USA). Freshly isolated epithelial cells and stromal cells were used for coculture and collection of condition medium as describe below. Some of the spare puri ed stromal cells (6000-8000 cells/cm 2 ) were plated into 100-mm petri dishes coated with bronectin (1 mg/ml, Invitrogen) and cultured in growth medium containing 10% FBS (Invitrogen), 1% antibiotics (Invitrogen) and 2 mmol/L glutamine (Invitrogen) in DMEM/F12 (Sigma-Aldrich, St Louis, MA, USA) for 7-14 days in a humidi ed carbon dioxide incubator at 37 o C in 5% CO 2 . The medium was changed every seven days until the cells reached 90% con uence.