IPC분류정보
국가/구분 |
United States(US) Patent
등록
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국제특허분류(IPC7판) |
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출원번호 |
US-0688031
(2000-10-10)
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발명자
/ 주소 |
- Funk, Walter D.
- Carpenter, Melissa K.
- Gold, Joseph D.
- Inokuma, Margaret S.
- Xu, Chunhui
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출원인 / 주소 |
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대리인 / 주소 |
Schiff, J. MichaelEarp, David J.
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인용정보 |
피인용 횟수 :
12 인용 특허 :
24 |
초록
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This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions prov
This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions provided in a suitable combination. Conditions that promote pPS cell growth without differentiation include supporting the culture on an extracellular matrix, and culturing the cells in a medium conditioned by another cell type. The cDNA libraries from such cultures are devoid of transcripts of feeder cell origin, relatively uncontaminated by transcripts from differentiated cells, and can have a high proportion of full-length transcripts. Subtraction libraries can also be produced that are enriched for transcripts modulated during differentiation.
대표청구항
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This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions prov
This disclosure provides a system for obtaining expression libraries from primate pluripotent stem (pPS) cells. pPS cells can be maintained in vitro without requiring a layer of feeder cells to inhibit differentiation. The role of the feeder cells is replaced by several other culture conditions provided in a suitable combination. Conditions that promote pPS cell growth without differentiation include supporting the culture on an extracellular matrix, and culturing the cells in a medium conditioned by another cell type. The cDNA libraries from such cultures are devoid of transcripts of feeder cell origin, relatively uncontaminated by transcripts from differentiated cells, and can have a high proportion of full-length transcripts. Subtraction libraries can also be produced that are enriched for transcripts modulated during differentiation. P>R7,and R8can each, independently, be selected from the group consisting of H, N3,R11,NO2,NHC(=NH)N(R10)2,NHC(O)R11,C(O)R11,Cl, Br, I, F, SR10,and (CH2)xC(=NH)N(R10)2where x is an integer from 0 to 3; wherein R9is: 1) OR10and R10is C(CH3)3,CH(CH3)2,CH2CH(CH3)2,CH(CH3)(CH2)mCH3where m is an integer from 0 to 3, or (CH2)mCH3,where m is an integer from 0 to 3, or 2) R9is selected from the group consisting of C(CH3)3,CH(CH3)2,CH2CH(CH3)2,CH(CH3)(CH2)mCH3where m is an integer from 0 to 3, CH2CH3,(CH2)2CH3,(CH2)3CH3,N(R10)2,NO2,NHC(=NH)N(R10)2,NHC(O)R11,C(O)R11,Cl, Br, I, F, SR10,and (CH2)xC(=NH)N(R10)2where x is an integer from 0 to 3; wherein R10=H, C(CH3)3,CH(CH3)2,CH2CH(CH3)2,CH(CH3)(CH2)mCH3where m is an integer from 0 to 3; or (CH2)mCH3,where m is an integer from 0 to 3; wherein R11=R10,OR10,or N(R10)2; wherein, R12=H or (CH2)n; where n is an integer from 0 to 3; or, also for General Structure I, wherein, R3can be selected from the group consisting of H, R11,OC(O)R11,NHC(O)R11,Cl, Br, l, F, CHO, C(O)R11,C(N--OH)NH2,OPO3R10,OPO2(CH2)jCH3,CH2PO3R10,OSO3R10,OSO2(CH2)jCH3,CH2SO3R10,or CN, where j is an integer from 0 to 3; R1and R5can each, independently, be selected from the group consisting of H, R11,OC(O)R11,NO2,NHC(O)R11,Cl, Br, I, F, CHO, C(O)R11,C(N--OH)NH2,OPO3R10,OPO2(CH2)jCH3,CH2PO3R10,OSO3R10,OSO2(CH2)jCH3,CH2SO3R10,or CN, where j is an integer from 0 to 3; wherein R2or R4=H, R11,OC(O)R11,NO2,NHC(O)R11,Cl, Br, I, F, CHO C(O)R11,C(N--OH)NH2,OPO3R10,OPO2(CH2)jCH3,CH2PO3R10,OSO3R10,OSO2(CH2)jCH3,CH2SO3R10,CN, where j is an integer from 0 to 3, (CR12=CR12)kCN, and (CR12=CR12)NO2,where k is an integer from 1 to 3 or wherein, R6,R7,and R8can each, independently, be selected from the group consisting of H, N3,R11,NO2,NHC(=NH)N(R10)2,NHC(O)R11,C(O)R11,Cl, Br, I, F, SR10,and (CH2)xC(=NH)N(R10)2where x is an integer from 0 to 3; wherein R9is: 1) OR10and R10is C(CH3)3,CH(CH3)2,
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