Epcam cd326 ab conjugated magnetic microbeads

Tumor (ccRCC) and normal kidney tissue specimens were immediately placed in a Petri dish with phosphate-buffered saline (PBS) 1x and cut into small pieces of about 1 mm³. Each small piece of tissue was placed on the surface of the Petri dish, previously wetted with 1ml of Dulbecco's Modified Eagle Medium (DMEM, Invitrogen) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and 1% penicillin-streptomycin-L-glutamine (Sigma-Aldrich). The Petri dish was maintained for 2 hours at 37°C in a humidified atmosphere with 5% CO₂. After this time, the small specimens were covered with 4 ml of DMEM and placed back in the incubator. Cell proliferation was obtained around the kidney specimens. Kidney epithelial tubular and neoplastic cells were isolated with EpCAM (CD326) Ab-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) under the effect of a magnetic field generated by the Mini MACS Separation Unit (Miltenyi Biotec), as previously described [35 (link)].

Tumor (ccRCC) and normal (N) kidney tissue specimens were immediately placed in a Petri dish with phosphate-buffered saline (PBS) 1× and cut into small pieces of about 1 mm³. Each small piece of tissue was placed on the surface of the Petri dish with Dulbecco’s Modified Eagle Medium (DMEM, Invitrogen, Life Technologies, Monza, Italy) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, Milan, Italy) and 1% penicillin-streptomycin-L-glutamine (Sigma-Aldrich, Milan, Italy). Cell proliferation was obtained around the kidney specimens as previously described [55 (link)]. Kidney epithelial tubular and neoplastic cells were isolated with EpCAM (CD326) Ab-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) under the effect of a magnetic field generated by the Mini MACS Separation Unit (Miltenyi Biotec), as previously described [55 (link)]. These cells were then characterized for EpCAM, CA IX and MUC1 by immunocytochemistry. Primary ccRCC cells were used for proliferation studies at the second passage.

Tumor (ccRCC) and normal (N) kidney tissue specimens were immediately placed in a Petri dish with phosphate-buffered saline (PBS) 1x and cut into small pieces of about 1 mm³. Each small piece of tissue was placed on the surface of the Petri dish with Dulbecco’s Modified Eagle Medium (DMEM, Invitrogen, Life Technologies, Monza, Italy) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich, Milan, Italy) and 1% penicillin-streptomycin-L-glutamine (Sigma-Aldrich, Milan, Italy). Cell proliferation was obtained around the kidney specimens as previously described [27 (link)]. Kidney epithelial tubular and neoplastic cells were isolated with EpCAM (CD326) Ab-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) under the effect of a magnetic field generated by the Mini MACS Separation Unit (Miltenyi Biotec), as previously described⁵. These cells were then characterized for EpCAM, CA IX, and NDUFA4L2 by immunocytochemistry. Primary ccRCC cells were used for proliferation studies at the second passage.