Stem cell prepn for treating tissue ischemia disease and its prepn process

A technology of stem cell preparation and ischemic disease, applied in the field of stem cell preparation and its preparation for the treatment of tissue ischemic disease

Inactive Publication Date: 2004-04-07
INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is not clear whether stem cells from human umbilical cord blood, peripheral blood and bone marrow can differentiate in vivo to form new blood vessels and improve tissue ischemia after transplantation, and there is no application of umbilical cord blood CD144 + Cell-derived endothelial progenitor cell transplantation for the treatment of ischemic diseases and a precedent for promoting angiogenesis with stem cell preparations in the treatment of ischemic diseases

Method used

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  • Stem cell prepn for treating tissue ischemia disease and its prepn process

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Embodiment approach 1

[0023] The preparation method of mononuclear cell stem cell preparation isolated from peripheral blood is as follows: use 300 units of recombinant human hematopoietic growth factor (rhG-CSF) to subcutaneously inject Bid to the patient to mobilize peripheral blood stem cells; use FACS CD45 to shoot CD45 / CD34 double fluorescent labeling method to monitor the number of CD34 positive cells in peripheral blood; on the fifth day, COBE Spectra Version 4 blood cell separator was used to separately collect and separate mononuclear cells; then purified and concentrated into a mononuclear cell suspension for transplantation; According to the number of cells, the excess cells were divided into 3 × 10 per bag 9 The number of cells is strictly in accordance with the cryopreservation procedure of stem cells and stored in liquid nitrogen for multiple use.

[0024] Clinical use method: The transplantation and process are performed in a sterile operating room with strict aseptic operation. Dou...

Embodiment approach 2

[0026] The preparation method of the CD133+ cell preparation isolated and extracted from the placenta and umbilical cord blood is: take the umbilical cord blood of normal delivery, and use CD133 + Magnetic Bead Separation Kit (MACS, Miltenyi Biotec, Germany) Immunomagnetic Bead Cell Sorting Isolation of CD133 from Cord Blood + Cells, the isolated CD133+ cells were placed in the cell culture medium for later use.

Embodiment approach 3

[0028] The preparation method of CD133+ cell-derived vascular endothelial stem cell preparation is: use magnetic bead affinity column (MACS) separation method to separate CD133+ cells, inoculate in the coated 4μg / cm 2 In the T25 culture flask of fibronectin (Fibronectin, Calbiochem Company); After adding 50ng / ml vascular endothelial growth factor (VEGF, Pepro Tech, UK), 20ng / ml interleukin 3 (IL-3, Kirin, Japan) and 50ng / ml Cultivate in the M199 (Gibco Company)+10%FBS (Hyclone Company) system of stem cell growth factor (SCF, Kirin) per ml for 2-4 weeks, and change the liquid twice a week; cell clusters and spindles can be seen after 4-7 days of culture adherent cells.

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Abstract

The present invention discloses one kind of stem cell preparation for treating tissue ischemia disease and its preparation process. The stem cell preparation includes mononuclear cell separated and extracted form peripheral blood, CD 133+ cell separated and extracted from other body's tissue, or vascular endothelial ancestral cell derived from CD133+cell. The preparation process includes separation and purification of stem cell, culture, preparation of cell suspension and other steps. The positive effect of the present invention is that the stem cell preparation has high curative effect and no toxic side effect to obliterative cardiac, cerebral and peripheral vascular and corresponding genetic defective diseases.

Description

technical field [0001] The invention relates to a stem cell preparation and a preparation method thereof, in particular to a stem cell preparation for treating tissue ischemic diseases and a preparation method thereof. Background technique [0002] As we all know, arteriosclerotic vascular occlusion and thrombotic ischemic disease are common and refractory diseases in contemporary society. Conventional treatment methods such as drug therapy, vascular bypass and interventional surgery are generally used for such diseases at present. The curative effect of drug therapy is not ideal, although the early effect of vascular bypass and interventional surgery is better, but the late effect is not significant, so the disability and mortality rate is high. [0003] In recent years, research on stem cell preparations has continued to deepen, and the unique curative effect of stem cell preparations on certain diseases has gradually attracted attention. Stem cells are the origin cells ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K35/51A61K35/545A61K45/00A61P7/00A61P7/02A61P9/10C12N5/078
Inventor 韩忠朝黄平平杨晨
Owner INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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