Q1. FibronectinÀ̶õ ¹«¾ùÀΰ¡¿ä?
FibronectinÀ̶õ ³ôÀº ºÐÀÚ·® (220-250 kD)À» °¡Áø ¼¼Æ÷¿Ü±âÁúÀÇ ±¸¼º ¿ä¼Ò Áß ÇϳªÀÔ´Ï´Ù. FibronectinÀº dimerÇüÅ·ΠÁ¸ÀçÇϸç, ¼¼Æ÷ÀÇ À̵¿(cell migration), ÇüÅÂ(morphology), »ýÀå(growth)°ú ºÎÂø(adhesion)¿¡ °ü¿©ÇÕ´Ï´Ù.
Q2. RetroNectinÀº ¾î¶»°Ô ¼¼Æ÷¿Í virus°£ co-localize Çϳª¿ä? Virus particleÀÌ °áÇÕÇÏ´Â À§Ä¡´Â ¾îµðÀΰ¡¿ä?
RetroNectinÀº ¾à 63 kDÀÇ ´Ü¹éÁú (574°³ÀÇ ¾Æ¹Ì³ë»êÀ¸·Î ±¸¼º)·Î central cell-binding domain (ÇüÅ III repeat 8, 9, 10)°ú high affinity heparin-binding domain II
(ÇüÅ III repeat 12, 13, 14)¿Í CS-1 site (human FibronectinÀÇ IIICS regionÀÇ ÀϺηΠ±¸¼ºµÇ¾î ÀÖ½À´Ï´Ù. RetroNectinÀº Lentivirus/Retrovirus particles°ú target ¼¼Æ÷¸¦
co-localization½ÃÅ´·Î½á, À¯ÀüÀÚ µµÀÔÈ¿À²À» Áõ°¡½Ãų ¼ö ÀÖ½À´Ï´Ù. Co-localizationÀº RetroNectin (CH-296)ÀÇ heparin-binding domain II°¡ Á÷Á¢ viral particle°ú °áÇÕÇÏ°í,
CS-1 site¿Í cell-binding domainÀÌ ¼¼Æ÷ÀÇ VLA-4, VLA-5°ú °áÇÕÇÏ¿© ÀÌ·ç¾îÁý´Ï´Ù.
Q3. Very Late Antigen-4 (VLA-4)¿Í Very Late Antigen-5 (VLA-5)´Â ¹«¾ùÀΰ¡¿ä?
VLA-4¿Í VLA-5´Â integrin superfamily¿¡ ¼ÓÇØ ÀÖÀ¸¸ç, integrin ¥á4¥â1°ú integrin ¥á5¥â1À¸·Î ¾Ë·ÁÁ® ÀÖ½À´Ï´Ù. IntegrinÀº ¼¼Æ÷ Ç¥¸éÀÇ heterodimer·Î,
¼¼Æ÷ ¿Ü ±âÁú°ú ¼¼Æ÷ ³» ¼¼Æ÷°ñ°Ý´Ü¹éÁúÀÇ »óÈ£ÀÛ¿ëÀ» Á¶ÀýÇÕ´Ï´Ù.
Q4. RetroNectinÀº ¾î¶² cell¿¡ »ç¿ëÇÒ ¼ö ÀÖ³ª¿ä?
RetroNectinÀº integrin VLA-4 ¶Ç´Â VLA-5¸¦ ¹ßÇöÇÏ´Â ¼¼Æ÷¿¡ ¸Å¿ì È¿°úÀûÀÔ´Ï´Ù. ÀÌ·¯ÇÑ integrin ºÐÀÚµéÀº FibronectinÀÇ CS1 site¿Í cell-binding domain¿¡ »óÈ£ÀÛ¿ëÇÕ´Ï´Ù.
Q5. ¾î¶² CellÀÌ VLA-4¿Í VLA-5¸¦ ¹ßÇöÇϳª¿ä?
VLA-4¸¦ ¹ßÇöÇÏ´Â ¼¼Æ÷´Â ´ëÇ¥ÀûÀ¸·Î, T cells, B cells, monocytes, NK cells, eosinophils, bone marrow monocytic cells°ú lymphoid progenitorsÀ̸ç, thymocytes,
activated T-cells°ú mast cellÀº VLA-5¸¦ ¹ßÇöÇÑ´Ù°í ¾Ë·ÁÁ® ÀÖ½À´Ï´Ù.
Q6. RetroNectinÀº ´Ù¾çÇÑ envelopes (Eco, Ampho, VSV-G pseudoÇüÅÂ)¿¡ »ç¿ë °¡´ÉÇϳª¿ä?
RetroNectinÀº ecotropic°ú amphotropic ¸ðµÎ È¿°úÀûÀ̸ç, GALV ÇüÅÂÀÇ envelope¿¡µµ »ç¿ë °¡´ÉÇÕ´Ï´Ù. VSV-G (Vesicular stomatitis virus G) pseudoÇüÅÂÀÇ Lentivirus/Retrovirusµµ
RetroNectinÀÌ ÄÚÆÃµÈ plate¸¦ »ç¿ëÇÏ¸é ¼¼Æ÷³» µµÀÔÈ¿À²À» ³ôÀÏ ¼ö ÀÖ½À´Ï´Ù.
Q7. RetroNectinÀº Lentivirus/RetrovirusÀÇ µµÀÔÈ¿À² Áõ°¡ ¸ñÀû ¿Ü¿¡ Viral vector ¶Ç´Â DNAÀÇ »ðÀÔ ¸ñÀûÀ¸·Î »ç¿ëÇÒ ¼ö ÀÖ³ª¿ä?
RetroNectinÀº lentivirusÀÇ µµÀÔ È¿À²À» Áõ°¡½ÃÅ°´Âµ¥ »ç¿ëÇÒ ¼ö ÀÖ½À´Ï´Ù. RetroNectinÀÌ Viral vector ¶Ç´Â DNAÀÇ µµÀÔ¿¡ È¿°úÀûÀÎÁö¿¡ ´ëÇؼ´Â ¾Ë·ÁÁ® ÀÖÁö ¾Ê½À´Ï´Ù.
Q8. RetroNectinÀ» »ç¿ëÇÏ¸é µµÀÔ È¿À²À» ¾ó¸¶³ª Áõ°¡½Ãų ¼ö ÀÖ³ª¿ä?
Viral supernatant infection method (¼¼Æ÷¿Í virus¸¦ ¼¯Àº ÈÄ RetroNectin¿¡ ³ëÃâ)¸¦ »ç¿ëÇÏ¿´À» °æ¿ì, RetroNectinÀº human CD34+ cell¿¡¼ 50~70%, mouse bone marrow mononuclear cell¿¡¼ 20~30%±îÁö virus µµÀÔÈ¿À²À» Áõ°¡½ÃÄ×½À´Ï´Ù.
Q9. RetroNectinÀ» »ç¿ëÇϸé ÀϹÝÀûÀ¸·Î ¼¼Æ÷ µ¶¼ºÀÌ ¾ó¸¶³ª °üÂûµÇ³ª¿ä?
Polybrene¿¡ ºñÇØ RetroNectinÀº µ¶¼ºÀÌ ¸Å¿ì ³·Àº °ÍÀ¸·Î È®ÀεǾú½À´Ï´Ù. ¿¹¸¦ µé¸é, 0.25 mg/§¢ÀÇ RetroNectinÀÌ Ã·°¡µÈ ¹èÁö¿¡¼ NIH3T3 cellsÀ» ¹è¾çÇÑ °á°ú, ¼¼Æ÷ÀÇ ¼ºÀå¿¡´Â °ÅÀÇ ¿µÇâÀÌ ¾ø¾ú½À´Ï´Ù.
Q10. RetroNectinÀº polybrene°ú ÇÔ²² »ç¿ëÇÒ ¼ö ÀÖ³ª¿ä?
RetroNectin°ú polybreneÀ» ÇÔ²² »ç¿ëÇÏ¸é ¿ÀÈ÷·Á À¯ÀüÀÚ µµÀÔÈ¿À²ÀÌ °¨¼ÒÇϹǷÎ, µ¿½Ã¿¡ »ç¿ëÇÏ´Â °ÍÀº ±ÇÀåµÇÁö ¾Ê½À´Ï´Ù.
Q11. RetroNectinÀº ¾î¶² ÇüÅÂÀÇ plate¸¦ »ç¿ëÇØ¾ß Çϳª¿ä?
RetroNectinÀÇ ÄÚÆÃÀ» À§Çؼ´Â ÄÚÆÃÀÌ µÇÁö ¾ÊÀº Á¶Á÷ ¹è¾ç plate³ª dish¸¦ »ç¿ëÇØ¾ß ÇÕ´Ï´Ù. ´ÙÄ«¶ó¹ÙÀÌ¿À¿¡¼´Â ÄÚÆà plate (¿¹: collagen ÄÚÆà plate)¿Í ÄÚÆÃÀÌ µÇÁö ¾ÊÀº plate¸¦ »ç¿ëÇÏ¿© ½ÇÇèÇÑ °á°ú, ÄÚÆà plate´Â È¿À²ÀÌ 1/5·Î °¨¼ÒÇÏ´Â °ÍÀ» È®ÀÎÇÏ¿´½À´Ï´Ù. ´ÙÄ«¶ó¹ÙÀÌ¿À¿¡¼´Â 6-well BD-Falcon (#351146) plateÀÇ »ç¿ëÀ» ±ÇÀåÇÕ´Ï´Ù.
Q12. ÄÚÆÃµÈ Á¶Á÷ ¹è¾ç plate¸¦ »ç¿ëÇÏ¿´À» ¶§ È¿À²Àº ¾î¶°ÇÑ°¡¿ä?
½ÇÇè °á°ú ÄÚÆÃÀÌ µÇÁö ¾ÊÀº plate¿¡ RetroNectinÀ» ÄÚÆÃÇÑ ÈÄ virus µµÀÔ È¿À² ÃøÁ¤°á°ú 30% ÀÌ»óÀ» º¸¿´Áö¸¸, ÄÚÆÃÀÌ µÇ¾îÀÖ´Â plate¿¡ RetroNectinÀ» ÄÚÆÃÇÏ¿© ÃøÁ¤ÇÑ
°á°ú 1~3% Á¤µµÀÇ È¿À²À» ³ªÅ¸³»¾ú½À´Ï´Ù.
¸¸¾à double ÄÚÆÃµÈ plate°¡ ÇÊ¿äÇÒ °æ¿ì, µÎ °¡Áö ¹æ¹ý¿¡ µû¶ó Å×½ºÆ®Çغ¸½Ã±â ¹Ù¶ø´Ï´Ù.
(1) RetroNectinÀ» ¸ÕÀú ÄÚÆÃÇÑ ÈÄ collagen (¶Ç´Â ´Ù¸¥ ¹°Áú) ÄÚÆÃ
(2) Collagen (¶Ç´Â ´Ù¸¥ ¹°Áú)À» ¸ÕÀú ÄÚÆÃÇÑ ÈÄ, RetroNectin ÄÚÆÃ
À§ÀÇ µÎ °¡Áö ¹æ¹ýÀÇ µµÀÔ È¿À²ÀÌ ÁÁÁö ¾Ê´Ù¸é, ¾Æ·¡ÀÇ ¹æ¹ýÀ¸·Î ÁøÇàÇØ Áֽñ⠹ٶø´Ï´Ù.
ÄÚÆõÇÁö ¾ÊÀº plate¿¡ RetroNectinÀ» ÄÚÆýÃÄÑ À¯ÀüÀÚ µµÀÔÀ» ÇÏ´Â ½ÇÇè ¹æ¹ýÀ» ±×´ë·Î ¼öÇàÇÕ´Ï´Ù. Virus µµÀÔ ÈÄ, 37¡É, 5% CO2 Á¶°Ç¿¡¼ 4½Ã°£ µ¿¾È ¹è¾çÇÕ´Ï´Ù.
±×¸®°í ¼¼Æ÷¸¦ ¸ð¾Æ¼ collagen (¶Ç´Â ´Ù¸¥ ¹°Áú)¿¡ ÄÚÆÃµÈ plate¿¡ 2~3ÀÏ°£ ¹è¾çÇÕ´Ï´Ù.
Q13. RetroNectinÀº ¾ó¸¶³ª ¸¹ÀÌ Freeze / Thaw°¡ °¡´ÉÇÑ°¡¿ä?
RetroNectin solution (1 §·/§¢)Àº ÃÖ´ë 5¹øÀÇ freeze / thaw°¡ °¡´ÉÇÕ´Ï´Ù. Ãß°¡ÀûÀÎ freeze/thaw´Â ÇÇÇÏ°í, ¿À¿°¿¡ ³ëÃâµÇÁö ¾Êµµ·Ï ÁÖÀÇ ¹Ù¶ø´Ï´Ù.
RetroNectinÀº -20¡É¿¡¼ 1³â, 4¡É¿¡¼ ¾à 3ÁÖ°£ ¾ÈÁ¤ÇÕ´Ï´Ù.
Q14. RetroNectinÀÌ ÄÚÆÃµÈ plate (pre-coated RetroNectin plate)´Â ¾ó¸¶³ª ¿À·¡ ¾ÈÁ¤ÇÑ°¡¿ä?
RetroNectinÀÌ ÄÚÆÃµÈ plate´Â 4¡É¿¡¼ ¾à 1~2°³¿ù º¸°ü °¡´ÉÇÕ´Ï´Ù. (ÇÏÁö¸¸, °¡´ÉÇÏ¸é »¡¸® »ç¿ëÇÏ´Â °ÍÀ» ±ÇÀåÇÕ´Ï´Ù.)
-20¡É¿¡´Â º¸°üÇÏÁö ¸¶¼¼¿ä.
Q15. RetroNectinÀº »ç¿ë Àü filter ¸ê±Õ °úÁ¤ÀÌ ÇÊ¿ä ÇÑ°¡¿ä? ¸¸¾à ÇÊ¿äÇÏ´Ù¸é, ¾î¶² ŸÀÔÀÇ Filter¸¦ »ç¿ëÇØ¾ß Çϳª¿ä?
RetroNectinÀº ¸ê±ÕµÈ solution »óÅ·ΠÁ¦°øµË´Ï´Ù. µû¶ó¼, Filter°úÁ¤Àº ÇÊ¿äÇÏÁö ¾Ê½À´Ï´Ù.
¸¸¾à RetroNectinÀ» filterÇÑ´Ù¸é, Millipore SLGV013SL ¶Ç´Â SLGV004SLÀ» »ç¿ëÇÒ °ÍÀ» ±ÇÀåÇÕ´Ï´Ù.
Q16. Clinical-grade (GMP) RetroNectinµµ ÆǸÅÇϳª¿ä?
³×, GMP °¡À̵å¶óÀο¡ µû¶ó ³ôÀº Ä÷¸®Æ¼·Î »ý»êµÈ RetroNectin (GMP) grade (Code T202)À» ÆǸÅÇÏ°í ÀÖ½À´Ï´Ù. ¿¬±¸¿ë Á¦Ç°À¸·Î, ex vivo clinical application¿¡
Àû¿ë °¡´ÉÇÕ´Ï´Ù. (¿¬±¸ ¸ñÀûÀ¸·Î Á¦ÇѵǾî ÀÖ½À´Ï´Ù.)
Q17. RetroNectinÀ» X-fold bag°ú °°Àº cell-culture bag¿¡ Coating ÇÒ ¼ö ÀÖ³ª¿ä?
³×, X-Fold bagÀº RetroNectinÀ» »ç¿ëÇÏ¿© ÄÚÆà °¡´ÉÇÕ´Ï´Ù. ¾Æ·¡ÀÇ Àû¿ë ¿¹¸¦ Âü°í¹Ù¶ø´Ï´Ù.
For X-Fold bag (85 §¯
2)
a) RetroNectin*À» 4¡É¿¡¼ overnight ¶Ç´Â »ó¿Â¿¡¼ 2½Ã°£ µ¿¾È ÄÚÆÃÇÕ´Ï´Ù.
*ÃÖ¼ÒÇÑ 9 §¢ÀÇ RetroNectin (20 ¥ìg/§¢)À» »ç¿ëÇϼ¼¿ä.
b) PBS·Î wash ÇÕ´Ï´Ù. (30 §¢ x 3¹ø)
c) Lentivirus/Retrovirus¸¦ ÷°¡ÇÕ´Ï´Ù. (4~6½Ã°£ µ¿¾È)
d) PBS·Î wash ÇÕ´Ï´Ù. (30 §¢)
e) ¼¼Æ÷¸¦ ÷°¡ÇÕ´Ï´Ù. (1 x 104 cells/§¯2)
f) 37¡É, 5% CO2¿¡¼ 3ÀÏ°£ ¹è¾çÇÕ´Ï´Ù.
ÁÖÀÇ : ÀÌ ¹æ¹ýÀº ´Ù¸¥ ÇüÅÂ, ´Ù¸¥ ºê·£µåÀÇ cell culture bag°ú ȣȯµÇÁö ¾ÊÀ» ¼ö ÀÖ½À´Ï´Ù.
Q18. µµÀÔ È¿À²À» Áõ°¡½ÃÅ°´Â ¹æ¹ýÀÌ ÀÖÀ»±î¿ä?
´ÙÀ½ÀÇ ¹æ¹ýÀº RetroNectinÀ» »ç¿ëÇÏ¿© µµÀÔ È¿À²À» Áõ°¡½ÃÅ°´Â ¹æ¹ýÀÔ´Ï´Ù. ¾Æ·¡ÀÇ ¹æ¹ýÀ» »ç¿ëÇÏÁö ¾Ê´õ¶óµµ, RetroNectinÀº Fibronectin¿¡ ºñÇØ ³ôÀº È¿À²À» º¸ÀÔ´Ï´Ù.
- a) °í¿ª°¡ÀÇ(>106 cfu/§¢ ÀÌ»ó) Lentivirus/Retrovirus¸¦ ÁغñÇÕ´Ï´Ù. 60~70%ÀÇ µµÀÔ È¿À²¿¡ ¾ò±â À§Çؼ´Â Lentivirus/Retrovirus solution (¿¹¸¦ µé¾î, measured titer ¡Ã 1.15x 106)À» 10¹è ÀÌ»ó ³óÃà½ÃÅ°°í, ÀÌ°ÍÀ» pre-load ÇÕ´Ï´Ù.
- b) Pre-load Virus particleÀº RetroNectinÀÌ ÄÚÆÃµÈ plate¿¡ 200-250 §¡/§¯2À¸·Î ÷°¡ÇÏ°í, 4~5½Ã°£ µ¿¾È 32¡É¿¡¼ ¹è¾çÇÕ´Ï´Ù.
- c) Virus°¡ load µÈ plate°¡ ¸¶¸£Áö ¾Êµµ·Ï Á¶½ÉÇϸé¼, ºÎÀ¯¹°À» Á¦°ÅÇÏ°í, PBS·Î washÇÕ´Ï´Ù. PBS wash ÈÄ, Target cellÀ» Áï½Ã growth media¿Í ÇÔ²² Ãß°¡ ÇÕ´Ï´Ù. ¸¸¾à plate°¡ ¸¶¸£¸é, µµÀÔ È¿À²ÀÌ Å©°Ô °¨¼ÒµË´Ï´Ù.
Q19. RetroNectinÀÌ ÄÚÆÃµÈ plate·ÎºÎÅÍ ¼¼Æ÷¸¦ ºÐ¸®(dissociation) ÇÒ ¶§ ÃßõµÇ´Â ¹æ¹ýÀÌ ÀÖ³ª¿ä?
Fibroblast¿Í °°ÀÌ °ÇÏ°Ô ºÎÂøµÇ¾î ÀÖ´Â ¼¼Æ÷´Â Trypsin EDTA/phosphate-buffered saline (PBS) (without Ca
2+, Mg
2+)¸¦ »ç¿ëÇÏ¿© ºÐ¸®½Ãŵ´Ï´Ù.
¾àÇÏ°Ô ºÎÂøµÇ¾î ÀÖ´Â (¶Ç´Â ºÎÀ¯ »óÅÂ) ¼¼Æ÷´Â Trypsinº¸´Ù 0.02% EDTA/PBS solutionÀ» »ç¿ëÇÏ¿© ¼¼Æ÷¸¦ ºÐ¸®½Ãŵ´Ï´Ù. EDTA/PBS solutionÀ¸·Î ¼¼Æ÷°¡ ºÐ¸®µÇÁö ¾Ê¾Æ,
TrypsinÀ» »ç¿ëÇÏ´Â °ÍÀº ¼¼Æ÷¿¡°Ô damage¸¦ ÁÙ ¼ö ÀÖ½À´Ï´Ù.
¾àÇÏ°Ô ºÎÂøµÇ¾î ÀÖ´Â ¼¼Æ÷¸¦ À§ÇÑ ¹æ¹ý
- a) Transfection ÀÌÈÄ, plateÀÇ »óÃþ¾×À» centrifuge tube¿¡ ¸ð¾ÆµÓ´Ï´Ù.
- b) PBS·Î plate¸¦ washÇÏ¿© ºÎÀ¯ ¼¼Æ÷¸¦ ¸ð¾Æ µÓ´Ï´Ù.
- c) ºÎÂøµÇ¾î ÀÖ´Â ¼¼Æ÷¸¦ »ç¿ëÀÚ ¸Å´º¾ó¿¡ µû¶ó Cell Dissociation Buffer (GIBCO; enzyme free, PBS-based)·Î ºÐ¸®ÇÕ´Ï´Ù.
- d) ȹµæÇÑ ¼¼Æ÷¸¦ ÇϳªÀÇ centrifuge tube¿¡ ¸ð¿ì°í, centrifugeÇÏ¿© ¼¼Æ÷¸¦ ħÀü½Ãŵ´Ï´Ù.
- e) Centrifugation ÈÄ HBSS/Hepes¸¦ »ç¿ëÇÏ¿© ¼¼Æ÷¸¦ µÎ ¹ø washÇÏ°í, HBSS/Hepes¿¡ ÀÖ´Â ºÎÀ¯ ¼¼Æ÷´Â ´ÙÀ½ ´Ü°è½ÇÇè¿¡ »ç¿ëÇÕ´Ï´Ù.
Q20. RetroNectinÀÌ ÄÚÆÃµÈ plate·ÎºÎÅÍ Trypsin ¶Ç´Â Cell Dissociation Buffer¸¦ »ç¿ëÇÏ¿© ¼¼Æ÷¸¦ ºÐ¸®ÇÑ ÈÄ, ´Ù½Ã °°Àº plate¿¡ ¼¼Æ÷°¡ ºÎÂøµÇ³ª¿ä? ¸¸¾à 96 well plate¿¡¼ FACS¸¦ ÇÑ´Ù¸é, ¼¼Æ÷¸¦ »õ·Î¿î plate·Î ¿Å°Ü¾ß Çϳª¿ä?
Trypsin ¶Ç´Â Cell dissociation buffer¸¦ »ç¿ëÇÏ¿© ºÐ¸®½ÃŲ ¼¼Æ÷¸¦ ´Ù¸¥ plate·Î ¿Å±âÁö ¾Ê´Â´Ù¸é, °°Àº plate¿¡ ´Ù½Ã ºÎÂøµÉ °ÍÀÔ´Ï´Ù. ´ÙÄ«¶ó¹ÙÀÌ¿À´Â FACS ½ÇÇèÀ» À§Çؼ´Â multi-channel pipetteÀ» »ç¿ëÇÏ¿© ´Ù¸¥ plate·Î ¿Å±æ °ÍÀ» ±ÇÀåÇÕ´Ï´Ù.
Q21. RetroNectinÀ» »ç¿ëÇÏ¿© virus°¡ µµÀÔµÈ ¼¼Æ÷¸¦ plate·ÎºÎÅÍ resuspensionÇÏ´Â ¹æ¹ýÀÌ ÀÖ³ª¿ä?
¿¹) Jurkat cellÀÇ ¼¼Æ÷ Ç¥¸é ¸¶Ä¿ÀÎ CD154¿Í CD69¸¦ ¿°»öÇÏ¿©, Flow cytometry·Î ºÐ¼®ÇÏ°í ½ÍÀºµ¥, TrypsinÀ» »ç¿ëÇÏ¸é ¼¼Æ÷ Ç¥¸é ¸¶Ä¿°¡ degradation µÈ´Ù°í ÇÕ´Ï´Ù.
HL60°ú K562¿Í °°Àº cell lineÀº ÆÄÀÌÆêÆÃÀ¸·Î ½±°Ô ¸ðÀÌ´Â °ÍÀ» È®ÀÎÇÏ¿´½À´Ï´Ù. ÇÏÁö¸¸, ÀϺΠcell lineÀÇ °æ¿ì trypsinÀ» »ç¿ëÇÏÁö ¾ÊÀ¸¸é ¸ðÀ¸´Â °ÍÀÌ ¾î·Æ½À´Ï´Ù.
Jurkat cellÀ» resuspend ÇÏ´Â °ÍÀº ´ÙÄ«¶ó¹ÙÀÌ¿À¿¡¼ È®ÀεÇÁö ¾Ê¾Ò½À´Ï´Ù. ÀϹÝÀûÀ¸·Î, pipettingÀ¸·Î ¼¼Æ÷¸¦ ¸ðÀ¸´Â °ÍÀÌ ¾î·Æ´Ù¸é, µµÀÔ 24½Ã°£ ÀÌÈÄ trypsinÀ» ó¸®ÇÏ¿©
¼¼Æ÷¸¦ ¸ðÀ¸°í, »õ·Î¿î dish·Î ¿Å±â´Â °ÍÀ» ÃßõÇÕ´Ï´Ù. ±×¸®°í, ÀÌƲ°£ ¼¼Æ÷¸¦ ºÎÀ¯ »óÅ·Π¹è¾çÇÑ ÈÄ, Flow cytometry¸¦ »ç¿ëÇÏ¿© ºÎÀ¯ ¼¼Æ÷¸¦ ¼öÁýÇÕ´Ï´Ù.