Methods and compositions for increased safety of stem cell-derived populations
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12N-005/0735
C12N-005/074
출원번호
US-0179175
(2016-06-10)
등록번호
US-10179901
(2019-01-15)
발명자
/ 주소
Colton, Clark K.
Millman, Jeffrey R.
출원인 / 주소
Massachusetts Institute of Technology
대리인 / 주소
Wolf, Greenfield & Sacks, P.C.
인용정보
피인용 횟수 :
0인용 특허 :
18
초록▼
The invention provides methods and compositions relating to differentiated cell populations that derive from pluripotent stem cells. The methods relate to reducing the number of residual stem cells present in such populations. The compositions include differentiated cell populations that contain red
The invention provides methods and compositions relating to differentiated cell populations that derive from pluripotent stem cells. The methods relate to reducing the number of residual stem cells present in such populations. The compositions include differentiated cell populations that contain reduced number of stem cells or that contain no stem cells. Pluripotent stem cells may be reduced in number and/or function through exposure to low oxygen levels.
대표청구항▼
1. A method for reducing residual pluripotent stem cells in an in vitro differentiated cell population, comprising culturing an in vitro differentiated cell population at an oxygen partial pressure that is less than 142 mmHg for at least 1 day and less than 10 days, andharvesting the differentiated
1. A method for reducing residual pluripotent stem cells in an in vitro differentiated cell population, comprising culturing an in vitro differentiated cell population at an oxygen partial pressure that is less than 142 mmHg for at least 1 day and less than 10 days, andharvesting the differentiated cell population after the culturing step;wherein the harvested cell population comprises less than 1 pluripotent stem cell per 103 cells,wherein 50% or more of the cells in the in vitro differentiated cell population are positive for at least one differentiation marker before the culturing step. 2. The method of claim 1, wherein the in vitro differentiated cell population comprises non-cardiomyocyte lineage cells. 3. The method of claim 1, wherein the in vitro differentiated cell population comprises pancreatic islet cells. 4. The method of claim 1, wherein the in vitro differentiated cell population is cultured for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days or 9 days. 5. The method of claim 1, wherein the in vitro differentiated cell population is derived from in vitro differentiation of an embryonic stem cell or an induced pluripotent stem cell. 6. The method of claim 1, wherein the oxygen partial pressure is less than 140 mmHg, less than 120 mmHg, less than 100 mmHg, less than 80 mmHg, less than 70 mmHg, less than 60 mmHg, less than 50 mmHg, 40 less than 40 mmHg, less than 30 mmHg, less than 20 mmHg, less than 10 mmHg or lower. 7. The method of claim 1, wherein the oxygen partial pressure is in the range of 100-140 mmHg. 8. The method of claim 1, wherein the oxygen partial pressure is in the range of 50-100 mmHg. 9. The method of claim 1, wherein the oxygen partial pressure is in the range of 10-50 mmHg. 10. The method of claim 1, wherein the oxygen partial pressure is in the range of 4-10 mmHg. 11. The method of claim 1, wherein the in vitro differentiated cell population is cultured in a culture vessel comprising an oxygen permeable membrane. 12. The method of claim 1, wherein the in vitro differentiated cell population is cultured in a culture vessel comprising a silicon rubber membrane. 13. The method of claim 1, further comprising measuring the residual pluripotent stem cells in the cultured differentiated cell population. 14. A differentiated cell population obtained according to the method of claim 1. 15. A method for reducing residual pluripotent stem cells in an in vitro differentiated cell population, comprising culturing an in vitro differentiated cell population at an oxygen partial pressure that is less than 142 mmHg for at least 1 day and less than 10 days to reduce the pluripotent stem cell number to less than 1 pluripotent stem cell per 105 cells, andharvesting the differentiated cell population after the culturing step. 16. A method for reducing residual pluripotent stem cells in an in vitro differentiated cell population, comprising differentiating a population of pluripotent stem cells into an in vitro differentiated cell population under normoxic conditions,culturing the in vitro differentiated cell population at an oxygen partial pressure that is less than 142 mmHg for at least 1 day and less than 10 days, andharvesting the differentiated cell population after the culturing step;wherein the harvested cell population comprises less than 1 pluripotent stem cell per 103 cells. 17. The method of claim 16, wherein 50% or more of the cells in the in vitro differentiated cell population are positive for at least one differentiation marker before the culturing step.
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