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Mixing stem cell injection and preparation method thereof

A technology of mixing stem cells and injections, which is applied in the direction of anti-toxins, drug combinations, and pharmaceutical formulas, can solve the problems of umbilical cord blood hematopoietic stem cells restricting the scope of clinical application, and achieve the goal of promoting proliferation and maintaining stem cell characteristics, good effect, and inhibiting agglutination. Effect

Active Publication Date: 2013-10-02
广州市天河诺亚生物工程有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Insufficient number of umbilical cord blood hematopoietic stem cells in the current umbilical cord blood stem cell injection restricts its clinical application

Method used

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  • Mixing stem cell injection and preparation method thereof
  • Mixing stem cell injection and preparation method thereof
  • Mixing stem cell injection and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Umbilical cord blood and umbilical cord samples were obtained from healthy pregnant women and infants. Hepatitis B, hepatitis C, syphilis, AIDS, cytomegalovirus, TORCH, mycoplasma, chlamydia, G-6PD and thalassemia were all negative. After the specimens were collected, they were transported back to the blood bank to maintain the transport conditions at 4-8°C. Prepare mixed stem cell injections as follows.

[0039] 1. Preparation of Umbilical Cord Mesenchymal Stem Cells

[0040] Mesenchymal stem cells are extracted from the umbilical cord tissue of healthy people, and the obtained mesenchymal stem cells are cultured and expanded in vitro. After the primary cells are collected, they are subcultured at a ratio of 1:3. When the cells cover about 80% of the density of the bottom of the bottle, they are digested and collected with trypsin, and subcultured at the same ratio, up to the third generation. Collect the cells of the 3rd generation. At this time, the collected cell...

Embodiment 2

[0073] With reference to the method of embodiment 1, obtain about 1 * 10 7 ~5×10 7 Hematopoietic stem / progenitor cells and 2×10 7 mesenchymal stem cells. The prepared hematopoietic stem / progenitor cells and mesenchymal stem cells were added to normal saline containing 5% AB umbilical cord plasma to make the concentration of the two kinds of stem cells respectively 5×10 5 / ml and 2×10 5 / ml, prepared as mixed stem cell injection.

Embodiment 3

[0075] The mixed stem cell injection prepared in Example 2 can be mixed with umbilical cord blood mononuclear cells separated by conventional methods. The number of cord blood mononuclear cells should not be less than 5×10 8 indivual.

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Abstract

The invention discloses a mixing stem cell injection and a preparation method thereof, and belongs to the field of cell preparation agents. The injection of the invention is mixture of mesenchymal stem cells from umbilical cord source and hematopoietic stem / progenitor cells from the umbilical cord blood source; the method of the mixing stem cell injection comprises the following steps: CD34+ hematopoietic stem cells are separated from the fresh umbilical cord blood and purified, and are subjected to mixed culture and amplification together with the mesenchymal stem cells from the umbilical cord source, during the amplification, the culture system is culture medium including AB umbilical cord blood plasma the volume fraction of which is 15 percent and the cell factor, 6*104 mesenchymal stem cells and CD34+ hematopoietic stem cells with the concentration of 5*104 / ml are inoculated into a culture flask and are cultured for about 10 days, and two kinds of cells are obtained at the same time, so that the mixed stem cell injection including 5*105 / ml of hematopoietic stem / progenitor cells and 2*105 / ml of mesenchymal stem cells and containing normal saline with the volume fraction of AB umbilical cord blood plasma as 15 percent is obtained.

Description

technical field [0001] The invention belongs to the field of cell preparations, in particular to a mixed stem cell injection and a preparation method thereof. Background technique [0002] Hematopoietic stem cells are one of the components of blood. They are the initial cells that generate various blood cells. They are also called hematopoietic pluripotent stem cells. They exist in bone marrow, embryonic liver, peripheral blood and umbilical cord blood. It has both a high capacity for self-renewal and the ability to further differentiate hematological progenitor cells. At present, hematopoietic stem cells have been widely used clinically. Hematopoietic stem cell transplantation can be used to treat a variety of blood system diseases and immune system diseases, including hematological malignancies (such as leukemia, multiple myeloma, myelodysplastic syndrome, lymphoma, etc.), hemoglobinopathy, bone marrow hematopoietic failure ( Such as aplastic anemia), congenital metaboli...

Claims

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Application Information

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IPC IPC(8): A61K35/44A61P35/02A61P37/06A61P37/02A61P39/00A61K35/14A61K35/28A61K35/51
Inventor 魏伟嵐山芮许超
Owner 广州市天河诺亚生物工程有限公司
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