RU2011110455A - Улучшенная клеточная композиция и способы ее получения - Google Patents
Улучшенная клеточная композиция и способы ее получения Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract 41
- 239000000203 mixture Substances 0.000 title claims abstract 37
- 239000000243 solution Substances 0.000 claims abstract 53
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 claims abstract 34
- 102000008100 Human Serum Albumin Human genes 0.000 claims abstract 26
- 108091006905 Human Serum Albumin Proteins 0.000 claims abstract 26
- 238000010790 dilution Methods 0.000 claims abstract 21
- 239000012895 dilution Substances 0.000 claims abstract 21
- 229940119744 dextran 40 Drugs 0.000 claims abstract 20
- 229920002307 Dextran Polymers 0.000 claims abstract 14
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical group CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract 13
- 238000007865 diluting Methods 0.000 claims abstract 11
- 238000001914 filtration Methods 0.000 claims abstract 6
- 239000002577 cryoprotective agent Substances 0.000 claims abstract 5
- 238000005138 cryopreservation Methods 0.000 claims abstract 4
- 210000004027 cell Anatomy 0.000 claims 83
- 239000000853 adhesive Substances 0.000 claims 20
- 230000001070 adhesive effect Effects 0.000 claims 20
- 230000003169 placental effect Effects 0.000 claims 20
- 102100031573 Hematopoietic progenitor cell antigen CD34 Human genes 0.000 claims 8
- 101000777663 Homo sapiens Hematopoietic progenitor cell antigen CD34 Proteins 0.000 claims 8
- 102000003729 Neprilysin Human genes 0.000 claims 8
- 108090000028 Neprilysin Proteins 0.000 claims 8
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 claims 4
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 claims 4
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 claims 4
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 claims 4
- 238000000386 microscopy Methods 0.000 claims 2
- 238000010257 thawing Methods 0.000 claims 2
- 239000006285 cell suspension Substances 0.000 claims 1
- 239000008194 pharmaceutical composition Substances 0.000 claims 1
- 210000004991 placental stem cell Anatomy 0.000 claims 1
- 210000000130 stem cell Anatomy 0.000 claims 1
- 238000007920 subcutaneous administration Methods 0.000 claims 1
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/48—Reproductive organs
- A61K35/50—Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
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- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
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- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0221—Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
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- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0605—Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
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Abstract
1. Способ получения композиции, содержащей клетки, который включает: ! (а) контактирование указанных клеток с раствором, включающим декстран и сывороточный альбумин человека (HSA), для получения раствора, содержащего клетки; ! (b) фильтрацию раствора, содержащего клетки; ! (с) если указанный раствор, содержащий клетки, включает более примерно 10±3×106 клеток на миллилитр, разбавление указанных клеток до содержания не более примерно 10±3×106 клеток на миллилитр раствором первого разбавления, включающим декстран; и ! (d) необязательно разбавление раствора, содержащего клетки, раствором второго разбавления, содержащим декстран, но не содержащим HSA, с получением композиции, содержащей клетки. ! 2. Способ по п.1, где указанные клетки подвергаются криоконсервации после стадии (с) перед стадией (d). ! 3. Способ по п.1, где указанный декстран в указанном растворе первого разбавления или указанном растворе второго разбавления представляет собой декстран 40. ! 4. Способ по п.1, где указанный декстран в указанном растворе первого разбавления и указанном растворе второго разбавления представляет собой декстран 40. ! 5. Способ по п.1, где указанный декстран в указанном растворе первого разбавления представляет собой 5,5% декстрана 40. ! 6. Способ по п.1, где указанный HSA в указанном растворе, включающем HSA, представляет собой 10% HSA. ! 7. Способ по п.1, где указанный раствор первого разбавления включает HSA. ! 8. Способ по п.7, где указанный HSA в указанном растворе первого разбавления представляет собой 10% HSA. ! 9. Способ по п.1, где указанный раствор первого разбавления включает криопротектор. ! 10. Способ по п.9, где указанный криопротектор представляет собой ДМСО. !
Claims (42)
1. Способ получения композиции, содержащей клетки, который включает:
(а) контактирование указанных клеток с раствором, включающим декстран и сывороточный альбумин человека (HSA), для получения раствора, содержащего клетки;
(b) фильтрацию раствора, содержащего клетки;
(с) если указанный раствор, содержащий клетки, включает более примерно 10±3×106 клеток на миллилитр, разбавление указанных клеток до содержания не более примерно 10±3×106 клеток на миллилитр раствором первого разбавления, включающим декстран; и
(d) необязательно разбавление раствора, содержащего клетки, раствором второго разбавления, содержащим декстран, но не содержащим HSA, с получением композиции, содержащей клетки.
2. Способ по п.1, где указанные клетки подвергаются криоконсервации после стадии (с) перед стадией (d).
3. Способ по п.1, где указанный декстран в указанном растворе первого разбавления или указанном растворе второго разбавления представляет собой декстран 40.
4. Способ по п.1, где указанный декстран в указанном растворе первого разбавления и указанном растворе второго разбавления представляет собой декстран 40.
5. Способ по п.1, где указанный декстран в указанном растворе первого разбавления представляет собой 5,5% декстрана 40.
6. Способ по п.1, где указанный HSA в указанном растворе, включающем HSA, представляет собой 10% HSA.
7. Способ по п.1, где указанный раствор первого разбавления включает HSA.
8. Способ по п.7, где указанный HSA в указанном растворе первого разбавления представляет собой 10% HSA.
9. Способ по п.1, где указанный раствор первого разбавления включает криопротектор.
10. Способ по п.9, где указанный криопротектор представляет собой ДМСО.
11. Способ по п.1, где указанный декстран 40 в указанном растворе второго разбавления представляет собой 10% декстрана 40.
12. Способ по п.1, где указанный раствор на стадии (а) включает криопротектор.
13. Способ по п.1, где указанная композиция, содержащая клетки, включает от примерно 7,5% до примерно 9% декстрана.
14. Способ по п.1, где указанная композиция, содержащая клетки, включает от примерно 1,0±3×106 клеток на миллилитр до примерно 5,0±1,5×106 клеток на миллилитр.
15. Способ по п.1, где указанные клетки представляют собой плацентарные клетки.
16. Способ по п.15, где указанные стволовые клетки представляют собой изолированные адгезивные плацентарные клетки человека.
17. Способ получения композиции, содержащей клетки, который включает:
(a) суспендирование множества изолированных адгезивных плацентарных клеток человека в растворе, включающем 5,5% декстрана 40, 10% HSA, для получения раствора, содержащего клетки;
(b) фильтрацию раствора, содержащего клетки, через 70 мкм фильтр;
(с) разбавление раствора, содержащего клетки, раствором, содержащим 5,5% декстрана 40, 10% HSA и 5% ДМСО, до содержания не более примерно 10±3×106 клеток/мл;
(d) криоконсервацию клеток;
(е) оттаивание клеток; и
(f) необязательно разбавление раствора, содержащего клетки, в соотношении от 1:1 до 1:11 10% декстраном 40 для получения указанной композиции, включающей клетки.
18. Способ получения композиции, содержащей клетки, который включает:
(а) центрифугирование множества изолированных адгезивных плацентарных клеток человека для сбора клеток;
(b) суспендирование клеток в 5,5% декстране 40;
(с) центрифугирование клеток для сбора клеток;
(d) повторное суспендирование клеток в 5,5% растворе декстрана 40, который включает 10% HSA, для получения раствора, содержащего клетки;
(е) фильтрацию раствора, содержащего клетки, через 70 мкм - 100 мкм фильтр;
(f) разбавление раствора, содержащего клетки, в растворе, включающем 5,5% декстрана 40, 10% HSA и 5% ДМСО, до содержания не более примерно 10±3×106 клеток/мл;
(g) криоконсервацию клеток;
(h) оттаивание клеток;
(i) необязательно разбавление раствора, содержащего клетки, в соотношении от 1:1 до 1:11 10% декстраном 40 для получения композиции, включающей клетки.
19. Способ получения композиции, содержащей изолированные адгезивные плацентарные клетки человека, который включает:
(а) получение множества изолированных адгезивных плацентарных клеток человека в растворе, включающем 5,5% декстрана 40 и 10% сывороточного альбумина человека (HSA), для получения раствора, включающего изолированные адгезивные плацентарные клетки человека;
(b) фильтрацию указанного раствора, включающего изолированные адгезивные плацентарные клетки человека через фильтр, который удаляет различимые невооруженным глазом клеточные скопления, для получения отфильтрованных изолированных адгезивных плацентарных клеток человека;
(с) разбавление указанных отфильтрованных изолированных адгезивных плацентарных клеток человека количеством раствора, включающего 5,5% декстрана 40, 10% HSA и 5% диметилсульфоксида (ДМСО), достаточным для достижения содержания указанных отфильтрованных изолированных адгезивных плацентарных клеток человека примерно 10±3×106 клеток на миллилитр;
(d) разбавление указанных изолированных адгезивных планетарных клеток человека в соотношении от 1:1 до 1:11 изолированные адгезивные плацентарные клетки человека:декстран 40 для получения указанной композиции.
20. Способ по п.1, где указанный фильтр представляет собой 70 мкм фильтр.
21. Способ по п.1, где указанный фильтр представляет собой 100 мкм фильтр.
22. Способ по любому из пп.1, 6, 7 или 8, где указанные клетки представляют собой изолированные адгезивные плацентарные клетки человека.
23. Способ по п.22, где указанные изолированные адгезивные плацентарные клетки человека представляют собой CD10+, CD34- и CD105+.
24. Способ по п.23, где указанные CD10+, CD34- и CD105+ клетки представляют собой CD200+.
25. Способ по п.24, где указанные CD10+, CD34-, CD105+ и CD200+ клетки представляют собой CD45- или CD90+.
26. Способ по п.25, где указанные CD10+, CD34-, CD105+, CD200+ клетки представляют собой CD45- и CD90+.
27. Композиция, полученная способом по любому из пп.1, 6, 7 или 8.
28. Способ получения композиции, содержащей изолированные адгезивные плацентарные клетки человека, который включает контактирование указанных изолированных адгезивных плацентарных клеток человека с раствором, включающим 5,5% декстрана и 10% HSA; фильтрацию изолированных адгезивных плацентарных клеток человека; и разбавление указанных изолированных адгезивных плацентарных клеток человека до содержания примерно 10±3×106 клеток на миллилитр раствором, включающим 10% декстрана 40 (мас./об.).
29. Композиция, содержащая множество изолированных адгезивных плацентарных клеток человека в растворе, включающем 10% декстрана 40, где указанная композиция включает от примерно 1,0±0,3×106 клеток на миллилитр до примерно 5,0±1,5×106клеток на миллилитр, и где указанная композиция не содержит макроскоплений клеток.
30. Композиция по п.29, где указанные плацентарные стволовые клетки представляют собой оттаянные криоконсервированные клетки.
31. Композиция по п.29, где указанная композиция помещена в пакет (мешок).
32. Композиция по п.29, где указанная композиция не включает макроскоплений клеток.
33. Композиция по п.29, где указанная композиция включает менее примерно 200 микроскоплений клеток на 106 клеток.
34. Композиция по п.29, где указанная композиция включает менее примерно 150 микроскоплений клеток на 106 клеток.
35. Композиция по п.29, где указанная композиция включает менее примерно 100 микроскоплений клеток на 106 клеток.
36. Композиция по п.29, где указанные изолированные адгезивные плацентарные клетки человека представляют собой CD10+, CD34- и CD105+.
37. Композиция по п.36, где указанные CD10+, CD34- и CD105+ клетки представляют собой CD200+.
38. Композиция по п.37, где указанные CD10+, CD34-, CD105+ и CD200+ клетки представляют собой CD45- или CD90+.
39. Композиция по п.37, где указанные CD10+, CD34-, CD105+ и CD200+ представляют собой CD45- и CD90+.
40. Композиция по любому из пп.27, 29-39, где указанная композиция представляет собой фармацевтическую композицию.
41. Способ по п.1 или 14, дополнительно включающий концентрирование композиции, включающей клетки, до примерно от 5×106 клеток на миллилитр до 1×108 клеток на миллилитр.
42. Способ по п.41, дополнительно включающий подкожное введение пациенту композиции, содержащей клетки.
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US9057708P | 2008-08-20 | 2008-08-20 | |
US61/090,577 | 2008-08-20 | ||
PCT/US2009/004740 WO2010021714A2 (en) | 2008-08-20 | 2009-08-20 | Improved cell composition and methods of making the same |
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RU2015134559A Division RU2662676C1 (ru) | 2008-08-20 | 2009-08-20 | Улучшенная клеточная композиция и способы ее получения |
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RU2011110455A true RU2011110455A (ru) | 2012-09-27 |
RU2563518C2 RU2563518C2 (ru) | 2015-09-20 |
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RU2015134559A RU2662676C1 (ru) | 2008-08-20 | 2009-08-20 | Улучшенная клеточная композиция и способы ее получения |
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