Method and kit for separating stem cells

A stem cell and cell technology, applied in the field of biomedicine, can solve the problems of high cost, high technical personnel requirements, and impure stem cells.

Active Publication Date: 2013-10-23
江苏元生细胞工程科技发展有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common problems of these methods are: 1) the obtained stem cells are not pure, 2) it is inconvenient to use, and the cost is high, 3) the requirements for technical equipment are high, and the requirements for technical personnel are high in operation.
[0004] Therefore, the current technology for isolating stem cells still needs to be improved

Method used

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  • Method and kit for separating stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 (General Materials and General Methods)

[0035] Reagent test kit:

[0036] Reagent A: Erythrocyte Separation Solution

[0037] 0.5-0.6% methylcellulose (MC) (Switzerland FLUN K company).

[0038] The average viscosity of methylcellulose is 4,000cP. First, prepare a 0.4-0.6% by weight methylcellulose solution with pre-cooled 4°C normal saline, place it at 4°C overnight, and then shake it repeatedly until the methylcellulose is completely dissolved . The obtained methylcellulose solution was autoclaved at 0.1 MPa for 15 minutes, cooled to 4° C. and shaken vigorously. Methylcellulose becomes colorless and transparent after being completely dissolved, pH 7-8, and stored at 4°C for later use.

[0039] Reagent B: Stem Cell Purification Solution I

[0040] HBSS solution (purchased from Invitrogen Company) or GBSS solution or MEM culture fluid

[0041] Reagent C: Stem Cell Purification Solution II

[0042] 8-15% by weight iohexol solution: a predetermined amou...

Embodiment 2

[0055] Reagent A: Erythrocyte Separation Solution

[0056] 20 ng / ml human basic fibroblast growth factor (hFGF2) and 20 ng / ml human stem cell growth factor-α (pro-stem cell growth factor) were added to 0.5% methylcellulose.

[0057] Reagent B: Stem Cell Purification Solution I

[0058] 20 ng / ml human basic fibroblast growth factor (hFGF2) and 20 ng / ml human stem cell growth factor-α (original stem cell growth factor) were added to HBSS solution (purchased from Invitrogen).

[0059] Reagent C: Stem Cell Purification Solution II

[0060] 13% Nycodenz solution: Nycodenz 13g, HBSS solution (invitrogen company) 100ml, stirring > 1 hour, add 20ng / ml human basic fibroblast growth factor (hFGF2) and 20ng / ml human stem cell growth factor-α (original stem cell growth factor ).

[0061] Reagent D: cell treatment solution

[0062] Add 20ng / ml human basic fibroblast growth factor (hFGF2) and 20ng / ml human stem cell growth factor-α (original stem cell growth factor) to the phosphate buf...

Embodiment 3

[0065] Reagent A: Erythrocyte Separation Solution

[0066] 20 ng / ml human basic fibroblast growth factor (hFGF2) and 20 ng / ml human stem cell growth factor-α (pro-stem cell growth factor) were added to 0.5% methylcellulose.

[0067] Reagent B: Stem Cell Purification Solution I

[0068] 20 ng / ml human basic fibroblast growth factor (hFGF2) and 20 ng / ml human stem cell growth factor-α (original stem cell growth factor) were added to HBSS solution (purchased from Invitrogen).

[0069] Reagent C: Stem Cell Purification Solution II

[0070] 13% Nycodenz solution: Nycodenz 13g, HBSS solution (invitrogen company) 100ml, stirring > 1 hour, add 20ng / ml human basic fibroblast growth factor (hFGF2) and 20ng / ml human stem cell growth factor-α (original stem cell growth factor ).

[0071] Reagent D: cell treatment solution

[0072] Add 20ng / ml human basic fibroblast growth factor (hFGF2) and 20ng / ml human stem cell growth factor-α (original stem cell growth factor) to normal saline.

...

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Abstract

The invention relates to a method and a kit for separating stem cells and provides a composition for separating stem cells. The composition comprises iohexol and balanced salt solution, wherein the iohexol is dissolved in the balanced salt solution. With the adoption of the composition, the stem cells can be effectively separated.

Description

technical field [0001] The present invention relates to the field of biomedical technology, in particular to methods and kits for isolating stem cells, more specifically, the present invention relates to the use of iohexol in the preparation of stem cell purification liquid, a composition for isolating stem cells, and a method for isolating stem cells , a kit for isolating stem cells and the use of the kit for isolating stem cells. Background technique [0002] Human bone marrow and umbilical cord blood contain a large number of hematopoietic cells, various stem cells and a large number of mature cells. All lines of stem cells and mature white blood cells are nucleated cells. Nucleated cells include a variety of stem cells, mononucleated / multinucleated cells, lymphocytes, and mesenchymal cells. [0003] The separation methods of bone marrow red blood cells and nucleated cells in vitro include machine centrifugation and sedimentation. The machine centrifugation method is w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0789
Inventor 张文炜杨光华
Owner 江苏元生细胞工程科技发展有限公司
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