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

Abstract

The invention discloses a kit for in-vitro separation of karyocytes and removal of erythrocytes and a use method thereof. The kit comprises a precipitation solution, a purification solution, a suspension and an erythrocyte lysis solution, wherein the precipitation solution, the purification solution, the suspension and the erythrocyte lysis solution 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. The suspension is normal saline. The erythrocyte lysis solution comprises ammonium chloride having a concentration of 7.47g / L and trihydroxymethylaminomethane having a concentration of 2.6g / L. The invention also discloses 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 method for separating and purifying stem cells from bone marrow, umbilical cord blood and peripheral blood in the prior art has disadvantages such as low stem cell viability and recovery rate, and cannot lyse red blood cells therein

[0004] In our experiments, we believe that the use of 6% hydroxyethyl starch combined with Ficoll for the separation of monocytes in blood improves the recovery rate of monocytes, but it is still mixed with red blood cells.