▎ 摘　　要

NOVELTY - Use of modified porous membrane is claimed in process of isolation, enrichment and in situ culture of circulating tumor cells. USE - The modified porous membrane is useful in process of isolation, enrichment and in situ culture of circulating tumor cells (claimed). ADVANTAGE - The modified porous membrane realizes efficient and non-destructive separation and enrichment of circulating tumor cells in whole blood samples, removes red blood cells through lysate in advance, can distinguish white blood cells with similar sizes through staining, and thus improves detection accuracy, has good biocompatibility, can be directly transferred to the medium for culture observation, and avoids loss or inactivation of cell during complex processing. DETAILED DESCRIPTION - INDEPENDENT CLAIMS are included for the following: method for preparing modified porous polydimethylsiloxane (PDMS) membrane, which involves (a) dissolving the amino-terminated PDMS monomer into n-heptane, stirring and dissolving, adding crosslinking agent and catalyst, stirring evenly and reacting to obtain casting solution,(b) filtering using large-pore membrane, as substrate, passing the casting solution through large-pore membrane substrate by vacuum filtration to obtain porous PDMS membrane and then drying and (c) dissolving carboxy-containing compound and ethyl(dimethylamino propyl)carbodiimide/N-hydroxysuccinimide (EDC/NHS) mixed solution in 2-(N-morpholino)ethanesulfonic acid (MES) buffer solution to obtain impregnation solution, adding the porous PDMS membrane to the impregnation solution for reaction and then washing several times using deionized water and phosphate buffer saline (PBS) buffer solution to obtain modified porous PDMS membrane; and combined method of separation and enrichment of circulating tumor cells and in situ culture involves arranging modified porous PDMS membrane in the isolation device, adding the whole blood sample and red blood cell lysate to the top of the modified porous PDMS membrane and mixing evenly the whole blood sample and the lysate, then providing negative pressure on the lower layer of the modified porous PDMS membrane for isolation operation, passing the whole blood sample through the isolation membrane, trapping circulating tumor cells on isolation membrane, removing the upper residual waste liquid after isolation operation, then collecting the modified porous PDMS film containing the enriched circulating tumor cell and adding to the culture medium for in situ in vitro culture at 37℃ in carbon dioxide atmosphere, where the composition of red blood cell lysate comprises 8 g ammonium chloride, 1 g potassium carbonate, 3.7 g edetate disodium, 10 ml hydrochloric acid and deionized water make up to 100 ml and the culture medium comprises Dulbecco's modified Eagle medium or Roswell Park memorial institute-1640 medium containing 5-10 %mass bovine serum.