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Kit for in-vitro separation purification of karyocytes and use method thereof

A technique for separating and purifying nucleated cells, applied in the field of separating and purifying nucleated cells and kits in vitro, which can solve the problems of low stem cell viability and recovery rate, varying numbers, and affecting the stable collection of mononuclear cells

Inactive Publication Date: 2012-10-24
BEIJING TSCELL BIOTECH
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  • Summary
  • Abstract
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  • Application Information

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Problems solved by technology

[0003] The methods of separating and purifying stem cells from bone marrow, umbilical cord blood and peripheral blood in the prior art have disadvantages such as low stem cell viability and recovery rate. Preliminary experiments in this study found that the direct separation method using Ficoll and Percoll takes a long time to operate, and different batches of Different Feoll or Pcrcoll, different blood samples, and the same operation for the same sample, the number of mononuclear cells collected is different, which affects the stable collection of mononuclear cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Preparation method of cell processing kit (bone marrow, umbilical cord blood, peripheral blood) S100

[0033] Preparation of precipitation solution: Dissolve 2.40 g of hydroxyethyl starch in 40 ml of normal saline with a concentration of 6%, put it in a water bath at 37°C, and start counting for 8 hours from the temperature of the water bath at 37°C. After 8 hours in the water bath at 37°C, remove from the water bath Take it out, store at room temperature for half an hour.

[0034] Preparation of the purified solution: 60ml of HISTOPAQUE1077 solution (polysucrose sodium diatrizoate), and counted for 8 hours since the temperature of the refrigerator was stabilized at 4°C. After 8 hours at 4°C, take it out of the refrigerator at 4°C, and store it at room temperature for half an hour.

[0035] Preparation of the suspension: Measure 20ml of normal saline with a graduated cylinder and save it.

[0036] Cell Processing Kit (Bone Marrow, Cord Blood, Peripheral Blood) S100 In...

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Abstract

The invention discloses a kit for in-vitro separation purification of karyocytes and a use method thereof. The kit comprises a precipitation solution, a purification solution and a suspension, wherein the precipitation solution, the purification solution and the suspension are packaged separately; the precipitation solution is composed of normal saline and hydroxyethyl starch dissolved in the normal saline and has a weight volume concentration of 6%; the purification solution is a mixture of polysucrose and sodium diatrizoate; and the suspension is normal saline. The invention also provides a use method of the kit.

Description

technical field [0001] The invention relates to a method and kit for separating and purifying nucleated cells in bone marrow, umbilical cord blood or peripheral blood in vitro. Background technique [0002] Stem cells are primitive cells with the potential of self-replication and multi-directional differentiation. They are the origin cells of the body and the primitive cells that form various tissues and organs of the human body. Stem cells can be isolated and purified from bone marrow, umbilical cord blood, and peripheral blood. The commonly used isolation methods for mononuclear cells are as follows: 6% hydroxyethyl starch sedimentation method, methylcellulose sedimentation method, Ficoll separation method, Percoll separation method method, 3% gelatin separation method, ammonium chloride lysed red blood cell method, direct centrifugation method, etc. The commonly used isolation method of monocytes is Ficoll or Percoll direct separation method. That is, the diluted blood ...

Claims

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Application Information

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IPC IPC(8): C12N5/0786
Inventor 迟明国蔡云松
Owner BEIJING TSCELL BIOTECH