Tracing method for medulla desmohemoblast stem cell

A bone marrow mesenchymal stem cell technology, applied in the field of bone marrow mesenchymal stem cell tracing, can solve the problems of cell source disputes, inappropriate human body replantation research, and execution

Inactive Publication Date: 2007-07-25
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of effective identification and tracking monitoring methods for the current research on transplanted cells in situ in vivo, the role and outcome of exogenous MSCs in the repair of osteochondral defects, and the newly formed tissue engineered bone / cartilage in vivo The source of cells in
[0003] The existing commonly used methods for identifying cells all need to kill the experimental animals within a certain period of time after transplantation, and perform immunohistochemical sectioning on the osteochondral tissue in vitro to analyze and identify the transplanted cells. These invasive methods cannot dynamically , Accurately observe the migration, distribution, proliferation and other life processes of the same transplanted cells in the living host, and it is not suitable for human replantation research

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1 Using SPIO to label MSCs, and using 0.2T MRI in vivo to track the distribution and migration of autologous subcutaneous transplantation magnetization-labeled MSCs

[0011] 1.1 Materials and equipment

[0012] Superparamagnetic iron oxide (SPIO): provided by the Nanomaterials Laboratory of the School of Chemical Engineering, Southwest University, with a core diameter of (80±5)nm and a concentration of 17.4mg / ml;

[0013] Protamine sulfate (Pro): product of Sigma Company, diluted with distilled water to 10mg / ml mother solution, stored at -20°C;

[0014] Experimental animals: 14 male Japanese white rabbits aged 2-3 months with a weight of 2-2.5 kg were selected and fed adaptively for 1 week, and were randomly divided into 3 groups, and both lower limbs were included in the experiment:

[0015] Group A (experimental group): Autologous subcutaneous transplantation of SPIO / BrdU double-labeled MSCs (n=6)

[0016] Group B (negative control group): Autologous subcuta...

Embodiment 2

[0062] Example 2 Experimental study on the repair of articular cartilage defects in rabbit knees by magnetically labeled MSCs traced by MRI

[0063] Purpose:

[0064] To explore the feasibility of using 1.5T MRI in vivo tracer magnetic labeling MSCs distribution and migration in the articular cavity of rabbit cartilage defect model, and to evaluate the effect of cartilage repair

[0065] 2.1. Materials and methods

[0066] Materials used are the same as in Example 1

[0067] Experimental animals and grouping: 24 2-3-month-old male Japanese white rabbits with a weight of 2-2.5 kg were selected and fed adaptively for 1 week. They were randomly divided into 3 groups, and both lower limbs were included in the experiment:

[0068] Group A (experimental group): Transplant SPIO-labeled MSCs+scaffold complex group (n=6)

[0069] Group B (negative control group): self-transplanted unlabeled MSCs+scaffold complex group (n=6)

[0070] Group C (blank control group): the knee joint on ...

Embodiment 3

[0108] experimental method

[0109] 3.1. Preparation of Magnetic Marking Fluid (Fe-Pro):

[0110] Dilute SPIO to 100 μg / ml, 50 μg / ml and 25 μg / ml with serum-free DMEM / F12; dilute Pro to 6 μg / ml with serum-free DMEM / F12, and then mix equal amounts of diluted SIPO and Pro to obtain The final concentration of the labeling solution was Pro 3 μg / ml, and SIPO was 50 μg / ml, 25 μg / ml and 12.5 μg / ml respectively.

[0111] 3.2. Magnetic labeling of MSCs:

[0112] When the third-generation MSCs grow to 85% of the bottom area of ​​the culture bottle, discard the original serum-containing culture solution, rinse twice with PBS, take the Fe-Pro labeling solution and replace it in equal volume, at 37°C, 5% CO2, 100% saturation Humidity incubator incubated overnight 12h.

[0113] 3.3. Identification of magnetically labeled MSCs

[0114] 1. Prussian blue staining of magnetically labeled cells: positive blue-stained particles in the cytoplasm are positive for Prussian blue staining, count p...

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Abstract

A tracing method for the intermedullary filling stem cells includes such steps as using transfectant to modify superparamagnetic iron oxide to make it carry positive charges, using it to mark the cultured intermedullary filling stem cells, inoculating said cells onto scaffold, transplanting it in the body of animal, and using NMR imaging instrument for tracing the cells in animal body.

Description

technical field [0001] The invention relates to a tracing method of bone marrow mesenchymal stem cells, which is used for studying the survival, distribution and migration of exogenous bone marrow mesenchymal stem cells in vivo. Background technique [0002] In recent years, bone marrow mesenchymal stem cells (bone-marrow-derived mesenchymalstem cells, MSCs) have been induced to express osteoblast phenotype and chondrocyte phenotype, and have good in vitro expansion ability, which has become the current bone / One of the best seed cells for cartilage tissue engineering, important progress has been made in the treatment of bone / cartilage defect diseases with MSCs transplantation in animals, and it has a clear effect on the improvement of bone / cartilage defects. However, due to the lack of effective identification and tracking monitoring methods for the current research on transplanted cells in situ in vivo, the role and outcome of exogenous MSCs in the repair of osteochondral ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61B5/055A61B5/00
Inventor 金旭红杨柳
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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