Method for separating and purifying hemopoietic stem cells derived from umbilical cord blood and placentas
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A hematopoietic stem cell, separation and purification technology, which is applied in the field of separation and purification of hematopoietic stem cells derived from umbilical cord blood and placenta, can solve the problem that the yield of hematopoietic stem cells fails to reach the ideal and other problems.
Active Publication Date: 2018-08-24
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...
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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] Among them, V 红 >30, add HES to the blo...
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 in 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 in the first embodiment.
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Abstract
The invention provides a method for separating and purifying hemopoietic stem cells derived from umbilical cord blood and placentas and relates to a method for separating and purifying stem cells, aiming at solving the problems that a conventional hemopoietic stem cell separation and purification method only pays attention to the added amount of HES (Hydroxyethyl Starch) according to the volume ofa blood sample, and does not consider the ratio change of red blood cells in different blood samples. According to the method provided by the invention, the quantity of the red blood cells in umbilical cord blood and placenta cell suspended liquid is detected through a blood analyzer, and the HES is added according to an accurate ratio; then an accurate emission amount of the red blood cells andblood plasma, and the weight of the purified hemopoietic stem cells are realized through formula calculation and a blood flow speed controller, so that accurate adding of cryopreservation liquid is realized and the yield of the hemopoietic stem cells is maximized; separation and purification operation has rules to follow so that a condition that the adding of the HES and the cryopreservation liquid and the emission of the red blood cell plasma are carried out according to experience of people is avoided, the recycling of the hemopoietic stem cells in the samples and quality safety of the hemopoietic stem cells after being cryopreserved are ensured, and quality deviation caused by the operation of people is avoided.
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. Both early placenta and umbilical cord blood are discarded as medical waste because the value of stem cells is not understood. 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 blood system diseases and immune system diseases, including mali...
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