Method for peritoneal metastatic cell detection and isolation thereof
Provided herein is a method for the detection and isolation of metastatic cells from a sample obtained from a subject. Also provided is a device comprising a microchip for use in the method. Also provided is a method of isolating the metastatic cells for culture and characterization. Provided herein is a kit for the method therein.
1 . A method for detecting peritoneal metastatic cells in a sample obtained from a subject comprising: (a) flowing the sample using a microfluidic device at an effective wall shear stress, wherein a channel bottom of the microfluidic device is coated with P-selectin, wherein the effective wall shear stress is about 0.1-0.15 dyne/cm 2 , and contacting peritoneal metastatic cells and peritoneal non-metastatic cells in the sample with P-selectin such that the peritoneal non-metastatic cells are all washed away, and the peritoneal metastatic cells selectively bind to the P-selectin; and (b) detecting selective binding of the peritoneal metastatic cells to the P-selectin.
2 . The method of claim 1 , wherein the peritoneal metastatic cells are from gastrointestinal cancer or gynecological cancer.
3 . The method of claim 2 , wherein the gynecological cancer is ovarian cancer, uterine cancer, cervical cancer or vaginal cancer.
4 . The method of claim 1 , wherein the gastrointestinal cancer is liver cancer, colon cancer, prostate cancer, bladder cancer and rectal cancer.
5 . The method of claim 1 , further comprising a step of isolating the peritoneal metastatic cells.
6 . The method of claim 1 , further comprising a step of culturing the peritoneal metastatic cells.
7 . The method of claim 1 , further comprising a step of characterizing the peritoneal metastatic cells.
8 . The method of claim 1 , wherein the effective wall shear stress is about 0.1 dyne/cm 2 .
9 . The method of claim 1 . wherein the microfluidic device is a microfluidic chip.
10 . The method of claim 1 , wherein the peritoneal metastatic cells are detected by a label.
11 . The method of claim 1 , wherein the P-selectin is a P-selectin-Fc recombinant protein.
12 . The method of claim 1 , wherein the channel bottom is prepared by incubating with 1 μg mL −1 selectin-Fc recombinant protein in PBS at 4° C. overnight.
13 . The method of claim 12 , further comprising a step of EDTA detachment, wherein the step of EDTA detachment comprises introducing the sample in a binding buffer into the channel bottom coated with P-selectin recombinant protein at a shear stress of 0.1 dynes/cm 2 for 1 min followed by perfusion with PBS for 1 min at the shear stress of 0.1 dynes/cm 2 .