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Method for rapidly filtering and screening medulla ossium substrate stem cell

A rapid filtration and stem cell technology, applied in the direction of bone/connective tissue cells, animal cells, sterilization methods, etc., can solve the problems of long time, expensive, difficult clinical use, etc., to achieve less interference, short time consumption, and benefit The effect of clinical use

Active Publication Date: 2012-07-25
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1) Although the separation method of flow cytometry separates cells quickly and has high purity, it is expensive, and it is difficult to separate trace cells with too low content, and it cannot handle a large number of cell samples, and it is also difficult to obtain a sterile cell suspension
2) The immunomagnetic bead separation method has a great impact on the activity of cells, and the recovery rate is only 60%-75%, which may lead to complete inactivation of cells. The same operation is more complicated and expensive
However, it takes a long time, involves in vitro culture of cells, and is difficult for clinical use.
4) Density gradient centrifugation is a commonly used method for concentrating and separating cells, but it often requires the use of a separation medium (such as Ficoll or Hypaque solution, etc.). Washing away the separation medium, maintaining the sterility of the entire operation requires special equipment, which is expensive

Method used

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  • Method for rapidly filtering and screening medulla ossium substrate stem cell
  • Method for rapidly filtering and screening medulla ossium substrate stem cell
  • Method for rapidly filtering and screening medulla ossium substrate stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] see Image 6 .

[0023] Image 6 There are the first one-way flap valve 1, the switch valve 2, the second one-way flap valve 3, the exhaust hole 4, the peristaltic pump 5, the filter box 6, the injection pump 8, the first right connecting pipe 11, the first Left connecting pipe 12 , second right connecting pipe 13 , second left connecting pipe 14 , first pipe 31 , second pipe 32 , filter screen 61 , movable cover 62 .

[0024] The working method of the present invention is as follows: open the switch valve 2, inject 250ml of bone marrow blood after anticoagulation treatment into the first pipeline 31 through the injection pump 8, close the switch valve 2, open the switch of the peristaltic pump 5, and the bone marrow blood respectively flows into the second right connecting pipe 13. Enter the filter box 6 for filtration, and then flow to the second pipeline 32. Under the action of the peristaltic pump 5 and the filter screen 61, the bone marrow stromal stem cells in 2...

Embodiment 2

[0028] see Figure 7 .

[0029] Figure 7 There are the first one-way flap valve 1, the switch valve 2, the second one-way flap valve 3, the exhaust hole 4, the peristaltic pump 5, the filter box 6, the injection pump 8, the first right connecting pipe 11, the first Left connecting pipe 12 , second right connecting pipe 13 , second left connecting pipe 14 , first pipe 31 , second pipe 32 , filter screen 61 , movable cover 62 .

[0030] The working method of the present invention is as follows: open the switch valve 2, inject 300ml of bone marrow blood after anticoagulation treatment into the first pipeline 31 through the injection pump 8, close the switch valve 2, open the switch of the peristaltic pump 5, and the bone marrow blood respectively flows into the second right connecting pipe 13. Enter the filter box 6 for filtration, and then flow to the second pipeline 32. Under the action of the peristaltic pump 5 and the filter screen 61, the bone marrow stromal stem cells in...

Embodiment 3

[0034] see Figure 8 , 9, 10.

[0035] Figure 8 There are filter recombiners 60, filters 61, bone marrow stromal stem cells 71, other bone marrow cells 72,

[0036] Figure 9 There are filter recombiners 60, filters 61, bone marrow stromal stem cells 71, other bone marrow cells 72,

[0037] Figure 10 There are filter complex 60 , filter mesh 61 , bone marrow stromal stem cells 71 , other bone marrow cells 72 , and stem cell-scaffold composite material 7 .

[0038] During the operation, 250ml of bone marrow blood was drawn for anticoagulation. Tricalcium phosphate is selected as the material of the filter screen 61, and the pore size of the filter hole can allow the bone marrow cells to pass through smoothly. The anticoagulated bone marrow blood is injected into the filter complex 60 for filtration ( Figure 8 ). Using the unique adhesion characteristics of bone marrow stromal stem cells, the bone marrow stromal stem cells 71 are selectively stored and enriched on the...

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Abstract

The invention relates to a filtering method, in particular to a method for quickly filtering and screening bone marrow stromal cells. The method adopts the technical proposal that the method for quickly filtering and screening the bone marrow stromal cells comprises the following steps: firstly, performing anticoagulant treatment on bone marrow blood to be used; secondly, filtering the bone marrow blood after anticoagulant treatment through a filtration recombiner; and thirdly, collecting stem cell-frame composite materials. The method has the advantages that the method quickly recombines thecells and a vector (namely a filter screen material) when filtering and screening the bone marrow stromal cells simultaneously, so as to integrate the two procedures of cell screening and cell-vectorcombination into a whole. The filtering method has short time consumption, does not need in vitro culture, can be simultaneously performed in an operation, and is more favorable for clinic use because the volume of the screened stromal cells is smaller than that of cell suspension finally obtained by density gradient centrifugation technology.

Description

technical field [0001] The invention relates to a filtering method, in particular to a rapid filtering and screening method for bone marrow stromal stem cells. Background technique [0002] The repair of bone defects often requires bone grafting or filling with bone substitute materials, and the current research focus of tissue engineering in this field is the scaffold carrier compounded with stem cells. Bone marrow stromal stem cells are isolated by adherent culture or density gradient centrifugation, and then expanded in vitro. Although it is possible to isolate bone marrow stromal stem cells in vitro and prepare a scaffold carrier with cells, it is still in the laboratory research stage and needs to be implanted. There are still many technical and ethical problems in the human body, and the culture cycle is long, easy to contaminate, and the cost is high, which affects the clinical application and promotion. [0003] The current cell screening techniques mainly include: ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08C12M1/12C12N5/0775
Inventor 干耀恺戴尅戎汤亭亭
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE