A method for separating and purifying hematopoietic stem cells derived from umbilical cord blood and placenta
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A technology for separation and purification of hematopoietic stem cells, applied in the field of separation and purification of hematopoietic stem cells derived from umbilical cord blood and placenta, can solve the problems of unsatisfactory yield of hematopoietic stem cells, and achieve the effects of reducing errors, ensuring quality and ensuring cell activity
Active Publication Date: 2021-10-08
天晴干细胞股份有限公司
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[0004] In the process of separating and extracting hematopoietic stem cells from umbilical cord blood and placenta, regardless of the manual separation method or the automatic separation method of the instrument, the yield of hematopoietic stem cells has not reached the ideal level, and the average collection of hematopoietic stem cells can only be about 85%. And depending on the amount of umbilical cord blood, different operators have different yields
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specific Embodiment approach 1
[0079] Specific embodiment one: a kind of method for separating and purifying derived from umbilical cord blood and placental hematopoietic stem cells of the present embodiment, it is carried out according to the following steps:
[0080] 1. Sampling and counting before preparation:
[0081] After taking the blood sample and shaking it for 3 times, put the sample on the counter to draw and count;
[0082] 2. Weighing and removing red blood cells:
[0083] The blood samples collected and counted above were weighed according to the following formula, and the red blood cells were removed:
[0084] Net weight of blood sample=total weight-101, net volume of blood sample=net weight of blood / 1.03; the total weight is the total weight of separation bag and blood sample;
[0085] The red blood cell removal operation in the blood sample is performed according to the following formula:
[0086] V 红 = HCT × net volume of blood sample;
[0087] where V 红 >30, add HES to the blood sam...
specific Embodiment approach 2
[0111] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the HCT is the HCT in the blood sample. Others are the same as the first embodiment.
specific Embodiment approach 3
[0112] Embodiment 3: This embodiment is different from Embodiment 1 in that: in step 4, the rotor of the centrifuge is trimmed by placing the rotor in the centrifuge, setting it at 50 g, and centrifuging for 8 min, which is completed. Others are the same as the first embodiment.
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Abstract
The invention relates to a method for separating and purifying hematopoietic stem cells derived from umbilical cord blood and placenta, which relates to a method for separating and purifying stem cells. The invention aims to solve the problem that the conventional hematopoietic stem cell separation and purification method only pays attention to the amount of HES added according to the volume of the blood sample, and does not consider the change of the ratio of red blood cells in different blood samples. The present invention detects the number of red blood cells in the umbilical cord blood and placental cell suspension through a blood analyzer, adds the amount of HES according to a precise ratio, and then realizes the accurate discharge of red blood cells and plasma and the weight of hematopoietic stem cells after purification through formula calculation and blood flow rate controller , realize the precise addition of cryopreservation solution, maximize the yield of hematopoietic stem cells, and follow rules for separation and purification operations, thereby avoiding the addition of HES and cryopreservation solution and the discharge of red blood cell plasma according to personnel experience, ensuring hematopoiesis in the sample The recovery of stem cells and the quality safety after cryopreservation avoid the occurrence of quality deviation caused by personnel operations.
Description
technical field [0001] The invention relates to a method for separating and purifying hematopoietic stem cells derived from umbilical cord blood and placenta. Background technique [0002] Umbilical cord blood and placenta contain a large number of hematopoietic stem cells. Early-stage placenta and umbilical cord blood are both discarded as medical waste because people do not understand the value of stem cells. Recent studies have found that umbilical cord blood and placenta contain hematopoietic stem and progenitor cells that can rebuild human hematopoietic and immune systems, and can be used for hematopoietic stem cell transplantation to treat various diseases. Umbilical cord blood and placenta have become important sources of hematopoietic stem cells, especially unrelated hematopoietic stem cells. It is also a very important human biological resource. [0003] Hematopoietic stem cells can be used to treat a variety of hematological and immune system diseases, including...
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