
[ÁÖÀÇ] µ¿°áµÈ ¼¼Æ÷ÁÖÀÇ Çص¿(Thawing) ½Ã, ¼¼Æ÷ »ýÁ¸À² Çâ»óÀ» À§ÇÏ¿© collagen-coated plate ¶Ç´Â flaks¸¦ »ç¿ëÇϽʽÿÀ.
ÃßÈÄ ¼¼Æ÷ ¹è¾ç ½Ã¿¡µµ ¼¼Æ÷ÁÖÀÇ ºÎÂøÀ²(adherence)ÀÌ ÁÁÁö ¾Ê´Ù¸é collagen-coated plate ¶Ç´Â flask¸¦ »ç¿ëÇÏ´Â °ÍÀ» ±ÇÀåÇÕ´Ï´Ù.
NOTE: We recommend using collagen-coated plates or flasks for efficient culturing of frozen stocks. Vessels coated with compounds other than
collagen may also provide suitable growth substrates (e.g. poly-L-lysine), but only collagen-coated plates (e.g. BD BioCoat Cellware, Collagen Type I) have been tested
at Clontech. Once recovered, the cells may be cultured directly on tissue culture plastic. However, if adherence is poor, we recommend using only collagen-coated vessels.
- °í¿ª°¡ÀÇ ·»Æ¼¹ÙÀÌ·¯½º Á¦ÀÛÀ» À§ÇØ Á¦ÀÛµÈ 293T Clone
- 293FT Cell LineÀÇ 6¹è, HEK293 Cell Line¿¡ ºñÇÏ¿© 30¹è ¿ª°¡°¡ ³ôÀº ¹ÙÀÌ·¯½º »ý»ê
- ´Ù¾çÇÑ Á¾·ùÀÇ ·»Æ¼¹ÙÀÌ·¯½º º¤ÅÍ¿¡ Àû¿ë °¡´É
- Lenti-X¢â Packaging single shots (Code 631275)¿Í
ÇÔ²² »ç¿ëÇϸé 10^8 IFUÀÇ ¹ÙÀÌ·¯½º¸¦ Á¦ÀÛ °¡´É (10cm plate ±âÁØ)
Á¦Ç°¼³¸í
Lenti-X¢â 293T Cell LineÀº °í¿ª°¡ÀÇ ·»Æ¼¹ÙÀÌ·¯½º¸¦ Á¦ÀÛ °¡´ÉÇÑ ·»Æ¼¹ÙÀÌ·¯½º ÆÐŰ¡¿ë ¼¼Æ÷ÁÖÀÌ´Ù. º» ¼¼Æ÷ÁÖ¸¦ ÀÌ¿ëÇϸé ÀϹÝÀûÀ¸·Î »ç¿ëÇÏ´Â 293FT Cell Line°ú
HEK293 Cell LineÀ» ÀÌ¿ëÇÏ¿´À» ¶§º¸´Ù ¾à 6¹è-30¹è ¿ª°¡°¡ ³ôÀº ¹ÙÀÌ·¯½º¸¦ ¾òÀ» ¼ö ÀÖ´Ù (±×¸² 1)

±×¸² 1. Lenti-X¢â 293T Cell LineÀÇ ¶Ù¾î³ ¹ÙÀÌ·¯½º »ý»ê ´É·Â
We used our 4th generation lentiviral packaging system and one of our pLVX lentiviral vectors to compare the virus production of the Lenti-X 293T
Cell Line to that of two other commonly used HEK 293-based cell lines. Lenti-X 293T cells clearly outperformed the other cell lines?producing over
6 times more virus than 293FT cells and up to 30 times more virus than the parental HEK 293 cell line.
½Ã³ÊÁö È¿°ú : ³ôÀº ¿ª°¡¸¦ À§ÇÑ ÇØ´ä
¾Æ·¡ÀÇ ±¸¼º¿ä¼Ò¸¦ 293T ¼¼Æ÷¿Í ÇÔ²² »ç¿ëÇϸé ClontechÀÇ Lenti-X System Vector »Ó¸¸ ¾Æ´Ï¶ó ´Ù¸¥ lentivirus vector¸¦ ÀÌ¿ëÇÒ ¶§µµ ÃÖ°í ¿ª°¡ÀÇ ¾ÈÀüÇÑ lentivirus¸¦ »ý»êÇÒ ¼ö ÀÖ´Ù.
- ÃÖÀûÈµÈ Packaging Mix - ClontechÀÇ 4¼¼´ë packaging mixÀÎ Lenti-X¢â Packaging Single Shots´Â °í¿ª°¡ÀÇ Lentivirus¸¦ °£´ÜÇÏ°í È¿À²ÀûÀ¸·Î Á¦ÀÛÇÒ ¼ö ÀÖµµ·Ï ÃÖÀûÈ µÇ¾î ÀÖ´Ù. °¢°¢ÀÇ packaging vector¿Í transfection ½Ã¾àÀÌ ¸ðµÎ ÃÖÀûÀÇ ºñÀ²·Î È¥ÇÕµÇ¾î µ¿°á°ÇÁ¶ »óÅ·ΠÁ¦°øµÇ¸ç, lentiviral expression vectorÀÇ Ã·°¡¸¸À¸·Î ¾ÆÁÖ ¼Õ½±°Ô packagingÀ» ÁøÇàÇÒ ¼ö ÀÖ´Ù.
- Tet System Transactivation - Tetracycline-responsive promoter element(TRE)ÀÇ Tet-Off transactivationÀ» ÅëÇÏ¿© packaging¿¡ ÇÊ¿äÇÑ ¸î °¡Áö ÁÖ¿ä ±¸¼º¿ä¼ÒµéÀ» °íµµ·Î ¹ßÇö½ÃŲ´Ù. HEK293 ±â¹ÝÀÇ cell lines¿¡¼ Tet System À¯µµ¹ßÇöÀº CMV promoter¿¡ ÀÇÇØ »ý»êµÇ´Â ´Ù¸¥ ½Ã½ºÅÛ¿¡¼ º¸´Ù ÈξÀ ³ôÀº ¹ßÇöÀ» À̲ø¾î³½´Ù (1).
- ÃÖÀûÈµÈ transfection - xfect´Â °íÈ¿À²ÀÇ ºñµ¶¼º transfectionÀ» °¡´ÉÇÏ°Ô Çϸç, lipid-based transfection ¹ýº¸´Ù ´õ ³ôÀº ¹ßÇöÀ» À¯µµÇÑ´Ù. Lentiphos HT´Â 293T packaging cells¿¡ ÃÖÀûȵǾî ÀÖÀ¸¸ç, 99% ÀÌ»óÀÇ transfection È¿À²À» ¾òÀ» ¼ö ÀÖ´Ù. ÃÖÀûÈµÈ pH¿Í buffer Á¶¼º, ¿°³óµµ, ¹ÝÀÀ ¿ë·®°ú ÇÔ²² °£´ÜÇÑ ½ÇÇè Á¶ÀÛ °úÁ¤Àº ÃÖÀûÀÇ transfectionÀ» º¸ÀåÇÑ´Ù.
±¸¼ºÇ° (ÀÚ¼¼ÇÑ ³»¿ëÀº COA¸¦ ÂüÁ¶Çϼ¼¿ä)
1ml Lenti-X 293T Cell Line (2.0 x 106 cells/tube)
º¸Á¸
Liquid nitogren (-196 ¡É)
¶Ç´Â Á¦Ç° ¼ö·É ÈÄ, ¹Ù·Î À¶ÇØ ¹× ¹è¾ç ±ÇÀå
Shipping
Dry ice (-80 ¡É)
References
Yin, D. X. et al. (1996) Anal. Biochem. 235:195-201.