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Method and device for separating single karyocyte

A nuclear cell, single technology, used in the device of peripheral blood mononuclear cells, the separation of umbilical cord blood, and the field of bone marrow, can solve the problem of high red blood cell mixing, and achieve the effect of low red blood cell mixing, high efficiency and remarkable effect.

Active Publication Date: 2009-10-21
SHENZHEN BEIKE BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] However, this solution still has the problem of high red blood cell contamination

Method used

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  • Method and device for separating single karyocyte
  • Method and device for separating single karyocyte
  • Method and device for separating single karyocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] quadruple bags such as figure 1 As shown, it consists of 4 quadruple bags No. 1-4, as shown in the attached picture figure 1 shown.

[0073] Among them, the No. 1 bag is the main bag, which is used to hold the mixed liquid of umbilical cord blood, bone marrow or peripheral blood and lymphocyte separation fluid, and as a centrifuge container. Among them, the umbilical cord blood, bone marrow or peripheral blood need to be centrifuged in advance to remove The upper platelet-rich plasma layer reduces the rejection rate of trypan blue and the amount of red blood cells mixed in; the No. 2 bag is an empty bag, used to collect the liquid of the target cell layer; the No. 3 and No. 4 bags are normal saline bags, prefilled with normal saline, mainly For the cells of interest after aseptic washing.

[0074] The capacity of the 4 bags No. 1-4 can be selected according to the size of the sample volume, and the corresponding sterile plastic bag can range from 30ml to 500ml. Prefe...

Embodiment 2

[0086] The sample collection bag is bone marrow, the volume ratio of hydroxyethyl starch solution and normal saline is 1:5, the volume ratio of pretreated umbilical cord blood, bone marrow or peripheral blood and lymphocyte separation medium is 1:1, centrifuged during pretreatment Centrifugal condition is 2000rpm, 22 DEG C for 5min, centrifugal condition in step (3) is 1000rpm, centrifugal 30min under 22 DEG C, centrifugal condition is 2200rpm, centrifugal 7min under 22 DEG C during washing, other steps and conditions are the same as embodiment 1.

Embodiment 3

[0088] The sample collection bag is peripheral blood. The pretreatment centrifugation conditions are 2500rpm, 22°C for 5min, the centrifugation conditions in step (3) are 1500rpm, 22°C for 30min, and the washing conditions are 2800rpm, 22°C for 7min. Other steps and conditions are with embodiment 1.

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Abstract

The invention provides a method and a device for separating a single karyocyte, in particular a method and a device for separating single karyocyte of cord blood, marrow and peripheral blood. The single karyocyte can be separated without a hundred grades purification environment, and the method and the device are not limited by sample numbers and volumes, and has high efficiency, simple structure and no pollution, thereby being applied to laboratory research and human body treatment, and being easy to produce in batch.

Description

technical field [0001] The invention provides a method and device for isolating mononuclear cells, especially a device for isolating umbilical cord blood, bone marrow and peripheral blood mononuclear cells Background technique [0002] Umbilical cord blood, bone marrow, and peripheral blood are the three major sources of hematopoietic stem cells. Hematopoietic stem cell transplantation has become one of the most effective measures for treating various blood diseases, certain malignant solid tumors, some genetic diseases, and severe immunodeficiency. Clinical application has proved that hematopoietic stem cell transplantation has a good effect on rebuilding or improving bone marrow hematopoietic function. Mononuclear cells (MNC) (including monocytes, lymphocytes, stem cells, etc.) are the active ingredients of hematopoietic stem cell transplantation. / Isolation rate of progenitor cells or mesenchymal stem cells, maintenance of cell viability, and success or failure of transp...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12M1/00
Inventor 胡祥叶圣勤李国喜李建斌邢培清杜广有程四国
Owner SHENZHEN BEIKE BIOTECH
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