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1. (WO2013123734) OPTIMIZED IN VITRO NUCLEATED CELL SEPARATION KIT AND APPLICATION METHOD

Pub. No.:    WO/2013/123734    International Application No.:    PCT/CN2012/075972
Publication Date: Fri Aug 30 01:59:59 CEST 2013 International Filing Date: Fri May 25 01:59:59 CEST 2012
IPC: C12N 5/071
Applicants: NINGXIA ZHONGLIANDA BIOPHYSICS CO., LTD.
宁夏中联达生物有限公司
WANG, Huailin
王怀林
Inventors: WANG, Huailin
王怀林
Title: OPTIMIZED IN VITRO NUCLEATED CELL SEPARATION KIT AND APPLICATION METHOD
Abstract:
An optimized in vitro nucleated cell separation kit comprising two reagents. Reagent 1 comprises an erythrocyte precipitant, sodium chloride, or potassium chloride; reagent 2 comprises sodium diatrizoate-polysucrose 400#, sodium diatrizoate-dextran, or Percoll, and has a density between 1.076 and 1.090. Application method: inserting 100 ml of human bone marrow blood, umbilical cord blood, or peripheral blood directly into 200 ml of reagent 1 liquid; standing for 10 to 30 minutes; collecting the supernatant; centrifuging for five minutes at a speed of 2,000 revolutions/minute (1,100xg); discarding the supernatant and placing the remaining into a centrifugal tube containing reagent 2 liquid; performing a density gradient centrifugation for 30 minutes at a controlled rotation speed of 1,500 revolutions/minute; obtaining the nucleated cell layer by suction, and centrifuging and cleaning one to three times; diluting with 50 ml of a cell maintenance liquid for future use; taking 100 μl to submit for examination or to self-examine.