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ÎÒÃÇÀÏʦµÄÒâ˼ÊÇ£º½«¸Éϸ°ûÖ±½ÓÅàÑøÔÚSTEM CELL µÄÒ»¿î¼×»ùÏËÎ¬ËØÅàÑø»ùÀÐγÉEB£¬ÔÚ¼Óϸ°ûÒò×Ó½øÐÐÓÕµ¼¡­¡­ÎÒÃÇʵÑéÊÒÊÇ×öÔìѪ·½ÃæµÄÑо¿¡£¿ÉÊÇ£¬ÎÒ¿´ÁËSTEM CELLµÄһЩָµ¼¡£Ö¸µ¼ºÜ¼òµ¥¡£´ó¸ÅÈçÏ£º
step 1 Prepare Cells
Process mouse cells by:
? ammonium chloride lysis
? density gradient separation
? progenitor cell enrichment with
EasySep®, StemSep®, SpinSep® or
FACS
Wash cells (e.g. in Iscove¡¯s MDM plus
2% FBS), then count and adjust cell
concentration.
step 2 Add Cells to MethoCult®
Add cells to MethoCult® and vortex.
Let tube stand to allow bubbles
to dissipate.
step 3 Plate and Incubate
Dispense cells into pre-tested culture dishes
using syringe and blunt-end needle.
Incubate mouse cells for 7-14 days in
humidified incubator at 37¡ãC and 5% CO2.
step 4 Count Colonies
Count and evaluate colony types using
inverted microscope and gridded scoring
dishes. Alternatively, individual colonies
may be plucked for routine staining, PCR,
or cytogenetic analysis.
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