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Donor type hemopoietic chimera and its prepn process

A chimera and artificial blood technology, applied in the field of biological cells, can solve the problem that human/animal chimeras cannot be used as a source of human blood cells, and achieve the effect of solving the problem of blood-borne diseases, avoiding complex requirements and high safety.

Inactive Publication Date: 2007-03-21
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

[0005] The purpose of the present invention is to provide a donor-type hematopoietic chimera and its preparation method, which solves the problem that the existing human / animal chimera cannot be used as a source of human blood cells

Method used

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  • Donor type hemopoietic chimera and its prepn process
  • Donor type hemopoietic chimera and its prepn process
  • Donor type hemopoietic chimera and its prepn process

Examples

Experimental program
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Effect test

Embodiment 1

[0052] As shown in Figure 1, the donor-type human / pig hematopoietic chimera was constructed by injecting human-derived hematopoietic factors

[0053] Step 11: Preparation of hHSC for transplantation by separating, purifying and expanding human mononuclear cells (hMNC)

[0054] A) Using human umbilical cord blood mononuclear cells to separate and purify hHSC;

[0055] The hHSCs used are mainly derived from umbilical cord blood mononuclear cells. Healthy, full-term mothers are selected, and the umbilical cord blood is collected after the fetus is delivered naturally or after caesarean section, and sodium citrate is used for anticoagulation. Anticoagulated umbilical cord blood was thoroughly mixed with an equal amount of RPMI-1640 culture medium, and the lymphocyte separation medium was used to separate mononuclear cells (MNC) according to MACS-CD34 + Immunomagnetic Beads Separation Kit Instructions Carry out magnetic labeling on MNC, the labeled cells pass through a sorting col...

Embodiment 2

[0076] Construction of donor-type human-pig hematopoietic chimera by transfection of human gene

[0077] Similar to Example 1, the difference is:

[0078] 21) Immediately transplant human bone marrow mesenchymal stem cells transfected with human hematopoietic factors such as human erythropoietin (hEPO) after transplanting human hematopoietic stem cells (hHSC) to the selected suckling pigs.

[0079] Wherein, human bone marrow mesenchymal stem cells transfected with human hematopoietic factor genes are prepared through the following steps:

[0080] Step 211) Cloning human-derived hematopoietic factor genes such as hEPO. Retrieve the gene sequence from the gene bank (GenBank), design amplification primers (introduce restriction sites BamH I and Xho I), perform RT-PCR, recover the amplified fragment, and insert it into the eukaryotic expression vector (pcDNA3) after the sequence is correct. middle.

[0081] Among them, RT-PCR is an existing technology that combines reverse tran...

Embodiment 3

[0086] Construction of human-pig hematopoietic chimeras using transgenic pigs

[0087] Similar to Example 1, the difference is:

[0088] 31) The suckling pigs selected for the construction of human / pig hematopoietic chimeras are transgenic pigs.

[0089] 32) For the screened transgenic pigs, there is no need to inject human-derived hematopoietic factors, such as rhEPO, etc. after hHSC transplantation.

[0090] The application of the donor hematopoietic human / pig hematopoietic chimera of the present invention is further described in detail below:

[0091] A) Application of the donor-type human / pig hematopoietic chimera of the present invention as providing human red blood cells

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Abstract

The present invention discloses donor type hemopoietic chimera and its preparation process. The chimera has end blood cell from differentiation and development of human stem cell and hemopoietic function from human hemopoietic stem cell. The preparation process of the chimera includes the following steps: preparing human hemopoietic stem cell for transplantation with single human nuclear cell; screening animal and performing treatment for transplantation; transplanting human hemopoietic stem cell to the screened animal; and treating the animal after transplantation to obtain human / animal hemopoietic chimera. The present invention provides ideal human blood source substitute.

Description

technical field [0001] The invention relates to the technical field of biological cells, in particular to a hematopoietic chimera, its preparation method and the application of blood substitutes. Background technique [0002] In recent years, the problem of insufficient blood sources has become more and more serious, and the problem of pathogens transmitted by blood transfusion has also become more and more prominent. The use of blood substitutes is the fundamental way to solve these two problems. In the literature: Crawford JH, Chacko BK, Patel RP. Regulation of vascular function by haemoglobin [J]. Biochemical Society symposium. 2004, 71: 135-42 pointed out that at present, there are better plasma substitutes for clinical use, However, the research on red blood cell substitutes has not made significant progress so far: CF4 preparations have been proven to only delay blood transfusion but not replace blood transfusion, and hemoglobin preparations (including hemoglobin gelat...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/28C12N15/85C12N15/12C12N5/0786C12N5/0789
Inventor 蒋天伦赵树铭府伟灵周来新刘真范娅涵解志杰
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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