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Home > ÀüÁ¦Ç°º¸±â > ¼¼Æ÷¤ýÀ¯ÀüÀÚ Ä¡·á ¿¬±¸ > Overview > RetroNectin¢ç application - Enhanced transduction of hematopoietic stem cells

RetroNectin¢ç application - Enhanced transduction of hematopoietic stem cells

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Hematopoietic stem cell (HSC)°ú ºÐÈ­µÈ ¼¼Æ÷·ù (¿¹, primary T cell, macrophage µî) ¿Í °°Àº ´ëºÎºÐÀÇ hematopoietic stem cellÀº virus¸¦ ÀÌ¿ëÇÑ À¯ÀüÀÚ Àü´ÞÀÌ ¾î·Æ±â ¶§¹®¿¡ ´Ù¾çÇÑ Ç÷¾×°ü·Ã Áúº´ÀÇ À¯ÀüÀÚ Ä¡·á¸¦ À§ÇÑ ¿¬±¸¿¡ °É¸²µ¹ÀÌ µÉ ¼ö ÀÖ´Ù.
Lentivirus, retrovirus¸¦ ÀÌ¿ëÇÑ hematopoietic stem cellÀÇ À¯ÀüÀÚ µµÀÔÈ¿À²Àº Ÿ°Ù ¼¼Æ÷ÀÇ Áֱ⠻óÅÂ, ¼¼Æ÷Ç¥¸éÀÇ receptor ¹ßÇö Á¤µµ µî ´Ù¾çÇÑ ¿äÀÎ

Enhanced virus-cell colocalization
RetroNectin¢çÀº human fibronectin protein¿¡¼­ À¯·¡ÇÑ 3°¡Áö µµ¸ÞÀÎÀ¸·Î ±¸¼ºµÇ¾î ÀÖ´Â polypeptideÀÌ´Ù. Virus ÀÔÀÚ°¡ RetroNectin¢çÀÇ heparin binding domain°ú °áÇÕÇÔÀ¸·Î½á virus ÀÔÀÚ¿Í Á¶Ç÷¸ð¼¼Æ÷°£ÀÇ À§Ä¡¸¦ ¹°¸®ÀûÀ¸·Î °¡±õ°Ô ÇÏ¿© transduction È¿À²À» ³ôÀÌ´Â ¿ø¸®·Î, Á¶Ç÷¸ð¼¼Æ÷´Â ÁÖ·Î ¼¼Æ÷ Ç¥¸éÀÇ integrin receptorÀÎ VLA-4¿Í RetroNectin¢çÀÇ CS-1 »çÀÌÆ®°¡ °áÇÕÇÑ´Ù.


±×¸² 1. Hypothesized mechanism of RetroNectin¢ç-mediated enhancement of transduction

High-efficiency transduction of hematopoietic cells
RetroNectin¢çÀº ¹°¸®ÀûÀ¸·Î ¼¼Æ÷¿Í ¹ÙÀÌ·¯½º ÀÔÀÚ¸¦ °¡±õ°Ô ÇÏ¿© ¼¼Æ÷µ¶¼ºÀº ÃÖ¼ÒÈ­Çϸç À¯ÀüÀÚÀÇ µµÀÔÈ¿À²À» ³ôÀ̹ǷÎ, ´Ù¾çÇÑ Á¶Ç÷¸ð¼¼Æ÷¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù.

Gene transfer efficiency for hematopoietic cells1

Cell type

Efficiency (%)

Human CD34+ CD38- BMC2

95.5

Human PBMC3

91.2

TF-1

97.9

SupT1

97.3

Jurkat

80.1

K-562

90.4

HL-60

86.1

Monkey CD34+ BMC

72.0

Monkey CD4+ T-cell

85.0


1 TransductionÀº RetroNectin¢ç Á¦Ç°¸Å´º¾ó¿¡ ±âÀçµÈ RetroNectin¢ç-Bound Virus (RBV) MethodÀ» »ç¿ëÇÔ.
ÀÚ¼¼ÇÑ ³»¿ëÀº Á¦Ç°¸Å´º¾ó (¸µÅ©) ÂüÁ¶.
2 Bone marrow cells.
3 Peripheral blood mononuclear cells.

Improved transduction compared with conventional methods
RetroNectin¢çÀ» ÀÌ¿ëÇÏ¸é ±âÁ¸ÀÇ PolybreneÀ» »ç¿ëÇÑ transduction ¹æ¹ý¿¡ ºñÇØ È¿À²ÀÌ ³·Àº ¼¼Æ÷ ŸÀÔ¿¡µµ À¯ÀüÀÚ µµÀÔÈ¿À²À» ³ôÀÏ ¼ö ÀÖ´Ù.
Human CD34+¼¼Æ÷¿¡¼­ RetroNectin¢çÀº transduction È¿À²À» 50~70% ³ôÀÏ ¼ö ÀÖÀ» »Ó¸¸ ¾Æ´Ï¶ó (±×¸² 1), ¼¼Æ÷µ¶¼ºµµ °ÅÀÇ ¾ø´Â °ÍÀ» È®ÀÎÇÏ¿´´Ù.


±×¸² 2. RetroNectin¢ç°ú PolybreneÀÇ retrovirus transduction È¿À² ºñ±³
Retrovirus µµÀÔÈ¿À²À» ºñ±³Çϱâ À§ÇØ, Polybrene°ú RetroNectin¢çÀÇ °¢ ¸Å´º¾ó¿¡ µû¶ó Retrovirus transductionÀ» ÁøÇàÇÏ¿© human CD34+ HSC¿¡ GFP À¯ÀüÀÚ¸¦ ¹ßÇö½ÃÅ°°í, flow cytometry·Î Çü±¤À» ÃøÁ¤ÇÏ¿´´Ù. GFP ¹ßÇö·®À» ±×·¡ÇÁÀÇ XÃà¿¡, GFP¸¦ ¹ßÇöÇÏ´Â CD34+ ¼¼Æ÷ÀÇ ºñÀ²À» %·Î Ç¥±âÇÏ¿´´Ù.
±× °á°ú, RetroNectin¢çÀ» ÀÌ¿ëÇÑ °æ¿ì Polybrene º¸´Ù transduction È¿À²ÀÌ 86¹è ³ôÀº °ÍÀ» È®ÀÎÇÏ¿´´Ù (°¢°¢ 25.8%, 0.3%).

ÀÌ¿Í °°ÀÌ ±âÁ¸¿¡ À¯ÀüÀÚ µµÀÔÈ¿À²ÀÌ ³·¾Æ ¾î·Á¿òÀ» °Þ¾ú´ø Á¶Ç÷¸ð ¼¼Æ÷ ¿¬±¸¿¡ RetroNectin¢çÀ» ÀÌ¿ëÇÔÀ¸·Î½á Á¶Ç÷¸ð¼¼Æ÷¿Í lentivirus ¶Ç´Â retrovirusÀÇ co-localizationÀ» ÃËÁøÇÏ¿© ¼¼Æ÷ µ¶¼º ¾øÀÌ ³ôÀº È¿À²·Î À¯ÀüÀÚ¸¦ µµÀÔÇÒ ¼ö ÀÖ´Ù.

Comparison of retroviral transduction of a K562 cell line using transduction enhancers from different companies


±×¸² 3. ´Ù¾çÇÑ transduction enhancerÀÇ retroviral transduction È¿À² ºñ±³
´Ù¾çÇÑ Transduction enhancer¸¦ ÀÌ¿ëÇÏ¿© K562 cell lineÀÇ retrovirus µµÀÔÈ¿À²À» ºñ±³ÇÑ °á°ú,
RetroNectin¢çÀ» ÀÌ¿ëÇÏ¿´À» ¶§, Polybrene, protamine ¹× Ÿ»ç human fibronectinº¸´Ù ¿ùµîÈ÷ ³ôÀº È¿À²·Î transduction µÇ´Â °ÍÀ» È®ÀÎÇÏ¿´´Ù.