Multi-enzyme system used for separating patient tumor tissue-originated primary cells, application method thereof, purpose thereof, and relative kit thereof
A primary cell, tumor tissue technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc. It is not easy to survive in vitro and other problems, to achieve the effect of being suitable for large-scale promotion and application, easy to survive, easy to cryopreservation and recovery
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Embodiment 1
[0029] Example 1 Isolation of liver cancer tissue-derived primary cells
[0030] The multi-enzyme system of patient tumor tissue-derived primary cell isolation kit: 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase, 0.01-0.1% (weight) trypsin , 0.02-0.2% (weight) neutral protease, 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0031] A three-step digestion and separation technique is used to digest and separate the primary cells derived from the patient's tumor tissue. first step digestion : use 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase; The second step of digestion : use 0.01-0.1% (weight) trypsin, 0.02-0.2% (weight) neutral protease; The third step of digestion : Use 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0032] Isolation of primary cells derived from liver cancer tissue: 97% purity, 94% culture survival rate, 75% recovery culture survival rate after cryopreservation.
[0033] T...
Embodiment 2
[0034] Example 2 Separation of lung cancer tissue-derived primary cells
[0035] The multi-enzyme system of patient tumor tissue-derived primary cell isolation kit: 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase, 0.01-0.1% (weight) trypsin , 0.02-0.2% (weight) neutral protease, 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0036] A three-step digestion and separation technique is used to digest and separate the primary cells derived from the patient's tumor tissue. The second step of digestion : use 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase; The second step of digestion : use 0.01-0.1% (weight) trypsin, 0.02-0.2% (weight) neutral protease; The third step of digestion : Use 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0037] Isolation of lung cancer tissue-derived primary cells: 92% purity, 90% culture survival rate, 71% recovery culture survival rate after cryopreservation.
[0038] ...
Embodiment 3
[0039] Example 3 Isolation of breast cancer tissue-derived primary cells
[0040] The multi-enzyme system of patient tumor tissue-derived primary cell isolation kit: 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase, 0.01-0.1% (weight) trypsin , 0.02-0.2% (weight) neutral protease, 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0041] A three-step digestion and separation technique is used to digest and separate the primary cells derived from the patient's tumor tissue. first step digestion : use 0.02-0.2% (weight) collagenase I-IV, 0.001-0.01% (weight) hyaluronidase; The second step of digestion : use 0.01-0.1% (weight) trypsin, 0.02-0.2% (weight) neutral protease; The third step of digestion : Use 0.001-0.01% (weight) deoxyribonuclease I, 5-50U / ml elastase.
[0042] Isolated breast cancer tissue-derived primary cells: 95% purity, 93% culture survival rate, 73% recovery culture survival rate after cryopreservation.
[0043] The biol...
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