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Culture solution for reinforcing expression of fetal liver hemopoietic stem cell CD34 and preparation thereof

A technology of hematopoietic stem cells and culture medium, applied in the field of bioengineering, can solve the problems of whether cells inherit and have the characteristics of original stem cells, and can overcome the uncertainty of cell differentiation, increase the number of purification, and the effect of purification is safe and reliable.

Inactive Publication Date: 2008-12-03
SHANGHAI TIANSHENG BIOTECHOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the uncertainty of neural stem cells in the process of directional development and differentiation, it is still uncertain whether the differentiated cells inherit and have the characteristics of the original stem cells, so there is a greater risk in clinical application

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] In this embodiment, the culture medium for enhancing the expression of fetal liver hematopoietic stem cells CD34 has a composition ratio of 1 liter (L) when the total weight is:

[0046] RPMI-1640 250ml;

[0047] Hanks 110ml;

[0048] Human stem cell growth factor 24μg;

[0049] Human stem cell growth factor 16ng;

[0050] Human granulocyte-macrophage colony-stimulating factor 17ng;

[0051] CD135 ligand protein 92ng;

[0052] Human erythropoietic factor 12ng;

[0053] Human epithelial growth factor 92ng;

[0054] Human fibroblast production factor-b 92ng;

[0055] MEM margin.

[0056] ① Take MEM, RPMI-1640, and Hanks according to the above proportions, mix them well, and stir them thoroughly to prepare the basal culture medium;

[0057] ② Take human stem cell growth factor-2, human stem cell growth factor, human granulocyte-macrophage colony-stimulating factor, CD135 ligand protein, human erythropoiesis factor, human epithelial cell growth factor, human fibrobl...

Embodiment 2

[0062] The composition ratio of the culture medium for enhancing the expression of CD34 in fetal liver hematopoietic stem cells in this embodiment is as follows when the total weight is 1L:

[0063] RPMI-1640 300ml;

[0064] Hanks 70ml;

[0065] Human Stem Cell Growth Factor-220μg;

[0066] Human stem cell growth factor 32ng;

[0067] Human granulocyte-macrophage colony-stimulating factor 33ng;

[0068] CD135 ligand protein 108ng;

[0069] Human erythropoietic factor 28ng;

[0070] Human epithelial growth factor 108ng;

[0071] Human fibroblast production factor-b 108ng;

[0072] MEM margin.

[0073] ① Take MEM, RPMI-1640, and Hanks according to the above proportions, mix them well, and stir them thoroughly to prepare the basal culture medium;

[0074] ② Take human stem cell growth factor-2, human stem cell growth factor, human granulocyte-macrophage colony-stimulating factor, CD135 ligand protein, human erythropoiesis factor, human epithelial cell growth factor, human...

Embodiment 3

[0079] The composition ratio of the culture medium for enhancing the expression of CD34 in fetal liver hematopoietic stem cells in this embodiment is as follows when the total weight is 1L:

[0080] RPMI-1640 301ml;

[0081] Hanks 98ml;

[0082] Human Stem Cell Growth Factor-2 13μg;

[0083] Human stem cell growth factor 25ng;

[0084] Human granulocyte-macrophage colony-stimulating factor 30ng;

[0085] CD135 ligand protein 101ng;

[0086] Human erythropoietic factor 21ng;

[0087] Human epithelial growth factor 101ng;

[0088] Human fibroblast production factor-b 101ng;

[0089] MEM margin.

[0090] ① Take MEM, RPMI-1640, and Hanks according to the above proportions, mix them well, and stir them thoroughly to prepare the basal culture medium;

[0091] ② Take human stem cell growth factor-2, human stem cell growth factor, human granulocyte-macrophage colony-stimulating factor, CD135 ligand protein, human erythropoiesis factor, human epithelial cell growth factor, huma...

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Abstract

The invention relates to a culture solution of strengthening fetal liver hematopoietic stem cell CD34 expression and a preparation method thereof and belongs to the biological engineering technology field. The components and the weight percentage of the culture solution are as follows: 56.5-64.7 percent of MEM; 29.4-30.4 percent of RPMI-1640; and 5.88-13.0 percent of Hanks etc. The preparation method comprises the following steps: (1)preparing the basic culture solution;(2) selecting again relative components according to the weight percentage, adding the same to the basic culture solution in sequence; agitating and dissolving; (3) purifying a mesenchymal cell, putting in a culture bottle, culturing, washing, mixing, continuously culturing, collecting supernatant fluid, filtering, centrifuging, removing all cell fragments and non-soluble matters, and obtaining a conditioned culture solution; (4) mixing the conditioned culture solution with MEM, utilizing sodium bicarbonate to adjust the mixed culture solution to pH of 7.40 to 7.45, filtering and obtaining the culture solution. The culture solution strengthens expression of the CD34 expression characteristics of the stem cell. The stem cell reaches an identifiable and purified range, the identified quantity of the stem cell is improved, the culture solution is safe and reliable, and the clinical application is satisfied.

Description

technical field [0001] The invention relates to a culture medium in the technical field of bioengineering and a preparation method thereof. In particular, it relates to a culture solution for enhancing the expression of CD34 in fetal liver hematopoietic stem cells and a preparation method thereof. Background technique [0002] Hematopoietic stem cells were proposed at the beginning of the last century and their clinical application has shown their therapeutic potential. After stem cell transplantation for many hematological diseases, the clinical symptoms of patients can be improved, and the survival period of patients can be significantly prolonged. The main sources of hematopoietic stem cells include bone marrow-derived hematopoietic stem cells, umbilical cord blood hematopoietic stem cells, and fetal liver hematopoietic stem cells. Due to the different sources of hematopoietic stem cells, the antigenicity of stem cells is also different. The main manifestations are: bone...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08C12P21/04C12N5/0789
CPCC12N2501/125C12N2501/115C12N2501/11C12N2501/22C12N5/0647C12N2501/599C12N2502/14C12N2501/14
Inventor 郝晓南赵珺章永泰
Owner SHANGHAI TIANSHENG BIOTECHOLOGY
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