Recipients are 6- to 10-week-old virgin female SCID-beige mice. Before transplantation, cells are resuspended as single cells in PBS and counted from DCIS.COM, SUM225, or primary human DCIS cells. A 30-gauge Hamilton syringe, 50-μl capacity, with a blunt-ended 1/2-inch needle is used to deliver the cells. The mice are anesthetized, and a Y-incision is made on the abdomen to allow the skin covering the inguinal mammary fat pads to be peeled back to expose the inguinal gland. The nipple of the inguinal gland is snipped so that the needle can be directly inserted through the nipple. Two microliters of cell-culture medium (with 0.1% trypan blue) containing cells at a concentration of 2,500 to 5,000 cells/μl are injected; the injected liquid can be visually detected in the duct. The skin flaps are repositioned normally and held together with wound clips. The primary human DCIS was chopped very finely by using a Teflon block and razor blade or scalpel followed by overnight enzymatic digestion in DMEM/F12 with antibiotics, supplemented with collagenase (1.0 mg/ml) and hyaluronidase (100 U/ml).
Animal and human experiments were conducted by following protocols approved by the Baylor College of Medicine Animal Care and Use and Human Subjects Committee. An informed consent was deemed not to be required by the Human Subjects Committee. [See Additional data file1 for a video demonstration of the intraductal method.]
Animal and human experiments were conducted by following protocols approved by the Baylor College of Medicine Animal Care and Use and Human Subjects Committee. An informed consent was deemed not to be required by the Human Subjects Committee. [See Additional data file
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