¡Ø º» Á¦Ç°Àº ´ÙÄ«¶ó¹ÙÀÌ¿À¿¡¼ Àΰ£ iPS ¼¼Æ÷ À¯·¡ microgliaÀÇ Á¦Á¶ ¹æ¹ý¿¡ °üÇÑ Àü¼¼°è µ¶Á¡ ¶óÀ̼±½º¸¦ Shionogi Pharmaceutical Co., Ltd. ·ÎºÎÅÍ Ãëµæ ÈÄ, Á¦Á¶ ÆÇ¸ÅÇϰí ÀÖ½À´Ï´Ù.
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Cellartis¢ç DEF-CS¢â 500 Culture System (Code Y30010)·Î ¹è¾çÇÑ
Cellartis¢ç human iPS cell line 12 (ChiPSC12) Kit (Code Y00285)·ÎºÎÅÍ ºÐȵǾú½À´Ï´Ù.
- Central Nervous System (CNS, ÁßÃ߽Űæ°è) ³» ¸é¿ª ¹ÝÀÀ ¿¬±¸¸¦ À§ÇÑ hiPSC À¯·¡ microglia
- Iba1, P2Y12¿Í °°Àº microglial markerÀÇ ³ôÀº ¹ßÇö
- MicrogliaÀÇ ±â´ÉÀû Ư¡ È®ÀÎ - Amyloid ¥â Ž½Ä ¹ÝÀÀ, lipopolysaccharide Àڱؿ¡ ÀÇÇÑ cytokine ¹æÃâ, ATP¿¡ ÀÇÇÑ ¼¼Æ÷ À̵¿
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Cellartis¢ç Microglia´Â Àΰ£ ¿ªºÐÈÁٱ⼼Æ÷ÀÎ ChiPSC12·ÎºÎÅÍ ºÐÈµÈ microglia-like cell·Î½á, yolk sac macrophage ¹ß»ý °úÁ¤À» in vitro »ó¿¡¼ ÀçÇöÇÏ¿´´Ù. º» Á¦Ç°Àº MicrogliaÀÇ ÁÖ¿ä markerÀÎ Iba1, P2Y12¸¦ ³ô°Ô ¹ßÇöÇϰí,
µ¹±â ¸ð¾çÀÇ Çüųª amyloid ¥â ޽ĴÉÀÌ È®ÀεǴ µî ½ÇÁ¦ »ýü ³» ¼¼Æ÷¿¡¼ º¸À̴ Ư¼ºÀ» º¸ÀδÙ. ¶ÇÇÑ, »ýü ³» microglia¿¡¼ º¸ÀÌ´Â ATP À¯µµ¿¡ ÀÇÇÑ migration ¼ºÁúÀ̳ª, lipopolysaccharide¿¡ ÀÇÇØ cytokineÀ» ¹æÃâÇÏ´Â ±â´ÉÀ» º¸À¯Çϰí ÀÖ´Ù.

±×¸² 1. Cellartis¢ç Microglia (from ChiPSC12) ¼¼Æ÷ »çÁø

±×¸² 2. Microglia marker (Iba1, P2Y12)ÀÇ ¸é¿ª ¿°»ö
°ÅÀÇ ¸ðµç ¼¼Æ÷¿¡¼ Microglia markerÀÎ Iba1, P2Y12 ¹ßÇöÀ» È®ÀÎÇÏ¿´´Ù. (Iba1 = green, P2Y12 = Red, Dapi (Nuclear) = blue)

±×¸² 3. Cellartis¢ç MicrogliaÀÇ Amyloid ¥â Ž½Ä´É È®ÀÎ
2Àϰ£ ¹è¾çÇÑ Cellartis¢ç Microglia¸¦ ÀÌ¿ëÇÏ¿©, Amyloid ¥â ޽ĴÉÀ» È®ÀÎÇÏ¿´´Ù. (Amyloid ¥â = yellow, Phalloidin = red, Dapi (Nuclear) = blue)

±×¸² 4. Lipopolysaccharide¿¡ ÀÇÇÑ cytokine ¹æÃâ ±â´É
2Àϰ£ ¹è¾çÇÑ Cellartis¢ç Microglia (from ChiPSC12)ÀÇ lipopolysaccharide¿¡ ÀÇÇÑ cytokine ¹æÃâ ±â´ÉÀ» Æò°¡ÇÏ¿´´Ù. °¢ ³óµµÀÇ Lipopolysaccharide¸¦ ¹èÁö¿¡ ÷°¡ÇÑ ÈÄ, IL-6, TNF-¥á¿Í IL-1¥âÀÇ ¹æÃâ·®À» ÃøÁ¤ÇÑ °á°ú, IL-6 ¹× TNF-¥áÀÇ ¹æÃâÀº
LPS ³óµµ ÀÇÁ¸ÀûÀ¸·Î È®ÀεǾúÀ¸¸ç, IL-1¥âÀÇ ¹æÃâÀº LPS 30ng / ml Á¸Àç ÇÏ¿¡¼ BzATP ³óµµ ÀÇÁ¸ÀûÀÓÀ» ³ªÅ¸³Â´Ù.

±×¸² 5. ATP À¯µµ¿¡ ÀÇÇÑ migration ´É·Â
Cellartis¢ç Microglia (from ChiPSC12)¸¦ ÇØµ¿ÇÑ Á÷ÈÄ ¼¼Æ÷¿¡ Calcein AM Çü±¤À» Ç¥ÁöÇÏ¿´´Ù. ÀÌ ÈÄ, Disposable Chemotaxis System (Neuro Probe HP, Code. 106-8)¸¦ ÀÌ¿ëÇØ ATP À¯µµ¿¡ ÀÇÇÑ migration ´É·ÂÀ» Æò°¡ÇÏ¿´´Ù.
Cellartis¢ç Microglia (from ChiPSC12)´Â 0.1mM ATP Á¸Àç ÇÏ¿¡¼ chemotaxis¸¦ º¸¿´´Ù.
ÀÚ¼¼ÇÑ ±¸¼ºÀº CoA¸¦ Âü°íÇϼ¼¿ä.