Kit for in-vitro separation purification of karyocytes and removal of erythrocytes and use method of the kit

Abstract

The invention discloses a kit for in-vitro separation purification of karyocytes and removal of erythrocytes. The kit comprises a precipitation solution, a purification solution, a suspension solution and an erythrocyte lysis solution, wherein the precipitation solution, the purification solution, the suspension solution and the erythrocyte lysis solution are separatedly packaged. 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. The suspension solution is normal saline. The erythrocyte lysis solution comprises a stock solution 1 and a stock solution 2, wherein the stock solution 1 and the stock solution 2 are separatedly packaged; the stock solution 1 comprises 10g / L of glucose, 0.6g / L of potassium dihydrogen phosphate, 3.58g / L of Na2HPO4.7H2O and 0.1g / L of a phenol red solution; and the stock solution 2 comprises 1.86g / L of CaCl2.2H2O, 4.0g / L of potassium chloride, 80g / L of sodium chloride, 1.04g / L of magnesium chloride and 2.0g / L of MgSO4.7H2O. 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 and capable of removing red blood cells. 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 separat...

AI Technical Summary

Problems solved by technology

[0003] The prior art methods for separating and purifying stem cells from bone marrow, umbilical cord blood, and peripheral blood have disadvantages such as low stem cell viability and recovery rate, and cannot completely remove red blood cells.