Flow kit for screening umbilical cord mesenchymal stem cells with strong migratory ability
A technology of mesenchymal stem cells and kits, which is applied in the field of flow cytometry kits for screening umbilical cord mesenchymal stem cells with strong migration ability, can solve the problems of increased number of sample tubes, sample loss, emission light leakage, etc., to reduce Requirements, safety assurance, consumption reduction effect
Active Publication Date: 2020-06-05
TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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Problems solved by technology
[0005] First of all, the currently recognized optimal detection system in the industry is that each tube contains 0.5mL with a density of 1*10 6 For cell samples, the number of detection tubes is determined by the type of surface markers to be detected; secondly, considering that when different fluorescent dyes are used for multi-color labeling, the emitted light will leak into other detection channels, it is necessary to use a single-color light control tube and FMO control tubes to obtain accurate experimental data; if a single-color dye is used for detection, due to the characteristics of umbilical cord mesenchymal stem cells, multiple positive surface markers need to be detected, which will also increase the number of sample tubes and cause sample loss
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[0026] The present invention will be described in further detail below in conjunction with specific examples, and the following examples are only descriptive, not limiting, and cannot limit the protection scope of the present invention with this.
[0027] The present invention uses microspheres with standard diameters of 15 μm and 30 μm to assist in setting the area where fluorescence signals need to be collected in the FSC-SSC map, which is called “Gate” in the industry; detection of CD166, CD86, and CD80 is added; PE- Positive Mixture of CD105, APC-CD73, FITC-CD90, PerCP-Cy5.5-CD166 and FITC-CD11b, PE-CD34, PE-CD45, PE-CD19, PE-CD80, PE-CD86, FITC-HLA-DR Negative Mixture; StandardMixture, Isotype Mixture, Positive Mixture, Negative Mixture formula; the experimental steps and data processing methods during detection, including but not limited to the drop relationship between various reagents and sample tubes, purity calculation methods, how to Set the signal acquisition area ...
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The invention relates to a flow detection kit for screening umbilical cord mesenchymal stem cells with strong migration ability. The kit consists of: standard diameter microspheres, PE-CD105, APC-CD73, FITC-CD90, PerCP- Cy5.5‑CD166, FITC‑CD11b, PE‑CD34, PE‑CD45, PE‑CD19, PE‑CD80, PE‑CD86, FITC‑HLA‑DR and corresponding isotype control antibodies. By setting the standard "Gate" and increasing the detection of CD166, the present invention directly correlates the test results with the migration ability and effectiveness of the cell preparation; the present invention ensures that multiple immune rejection-related cell surface markers such as CD80\CD86 are negative, thereby ensuring Personal safety of clinical patients; the invention reduces the consumption of cell samples in the process of detecting cell surface markers.
Description
technical field [0001] This patent is mainly used for the quality control of umbilical cord mesenchymal stem cell preparations, and can provide a method for evaluating cell migration ability for related research, especially a flow cytometry kit for screening umbilical cord mesenchymal stem cells with strong migration ability. Background technique [0002] Umbilical cord mesenchymal stem cells (UC-MSC, umbilical cord mesenchymal stem cells) are important members of the mesenchymal stem cell family and belong to pluripotent stem cells. Umbilical cord mesenchymal stem cells have high differentiation potential, can differentiate into cartilage, fat, endothelium and other directions, and have a certain role in immune regulation. , compared with bone marrow mesenchymal stem cells), it has the advantages of abundant sources and simple preparation. At present, UC-MSC preparations have shown good clinical application prospects in the treatment of bone and joint diseases, neurodegene...
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IPC IPC(8): C12Q1/02G01N15/14
CPCG01N15/14G01N33/5029G01N33/5073Y02A50/30
Inventor 张硕马洁
Owner TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD