38 results about "Megakaryocyte Progenitor Cells" patented technology

The invention relates to methods of obtaining compositions for generating multipotent hematopoietic stem progenitor cells comprising expansion of hematopoietic stem cells in the presence of HDACI and IDM. Methods of obtaining compositions enriched in hematopoietic megakaryocyte progenitor cells are also provided. Compositions enriched for stem cells and populations of cells obtained therefrom are also provided by the invention.

The invention discloses a method for inducing megakaryoblast and megakaryocyte in vitro and a special culture medium thereof. The special culture medium is a serum free medium containing 50ng/mL of SCF, 50ng/mL of TPO, 20ng/mL of IL-3 and 50ng/mL of IL-6. The method comprises the following steps of: 1) separating single karyocyte from umbilical cord blood by using the conventional Ficoll density gradient centrifugation method, wherein red blood cells in the umbilical cord blood are settled by 6 percent hydroxyethyl starch; and 2) culturing the single karyocyte in vitro for 4 to 14 days by using the special culture medium at 37 DEG C in the presence of 5 percent CO2 to obtain a mixture of the megakaryoblast and the megakaryocyte. The invention provides a source for supplementing the megakaryoblast and the megakaryocyte, and has the advantages of simple operation, low cost, high differentiation efficiency and the like. The method is about to play an important role in the field of medicaments, and has wide application prospect.

Various forms of c-mpl agonist antibodies are shown to influence the replication, differentiation or maturation of blood cells, especially megakaryocytes and megakaryocyte progenitor cells. Accordingly, these compounds may be used for treatment of thrombocytopenia.

The invention discloses a method for increasing the directional differentiation quantity of cord blood megakaryoblasts. The method comprises the step of carrying out directional differentiation on cord blood hematopoietic stem cells serving as specimen cells in a specific culture medium containing resveratrol. Due to the adoption of the specific culture medium containing resveratrol, the operation for directional differentiation of megakaryoblasts becomes simpler, more convenient, safer and more efficient. Resveratrol is added while cell factors are added in the directional differentiation culture process from the cord blood hematopoietic stem cells to the megakaryoblasts, so that the effect of increasing the quantity of the generated megakaryoblasts is achieved. The method is simple and convenient to operate and low in cost, and the obtained megakaryoblasts are high in safety.

The present invention relates to mainly one system of efficient culturing and directive inducing stem cell differentiation for culturing and proliferating megakaryocyte. The system has umbilical cord blood and marrow stem cell as seed cell, and the serum-free culture medium comprising TPO, IL-11 and heparin as the main component for megakaryocyte proliferation. The megakaryocyte preparation prepared based on the present invention contains great amount of macronucleus ancestral cells and mature megakaryocyte as well as CD34+ cells, and has the functions of treating thrombocytopenia and improving the blood-forming function after stem cell transplantation. The present invention provides novel efficient cell treating preparation for various thrombocytopenia and hemocyitopenia.

The invention relates to the field of cell culture, in particular to a separation method for megakaryocyte progenitors. According to the method, stimulation culturing is performed on single karyocytes obtained through separation through rhG-CSFs, so that the content of megakaryocytes in the cells is increased, the cells processed through stimulation culturing are screened with immunomagnetic beads, and the megakaryocyte progenitors are obtained. The megakaryocyte progenitors are isolated by adopting the method, the yield is higher, and the number of the megakaryocytes (the phenotype is CD41a<+>/CD61<+>) obtained through induced directional differentiation is larger. Experiments show that 20-100 mL of umbilical cord blood is separated through the method, the number of the obtained megakaryocyte progenitors can reach 2.75*10<6>, the quantity percentage of the megakaryocytes in the obtained cells is 5%-10%, and after 14 days of induced directional differentiation, the total number of the megakaryocytes can be amplified by 132.5+/-21.2 times, and the proportion of the megakaryocytes can reach 81%.

The invention relates to the field of cell culture, in particular to cell preservation liquid and application thereof in protecting megakaryocyte progenitor cell activity. The cell preservation liquid comprises water, folic acid, albumin, an astragalus extract and sodium chloride. The cell preservation liquid can provide corresponding nutrients for cells in the preservation process, so that the activity of the cells is maintained, and the cells are not likely to be gathered or died. More preservation liquid can be provided for preserving the megakaryocyte progenitor cells, the cell activity can be kept at about 90% within 24 h, the cell preservation liquid is obviously better than a control group in which normal saline or the mixture of normal saline and albumin is adopted, and it proves that the preservation liquid can achieve the effect of well protecting the megakaryocyte progenitor cell activity.

The invention discloses an in-vitro culture method of umbilical cord blood megakaryocyte progenitor cells. According to the method, a specific megakaryocyte progenitor cell medium is inoculated with umbilical blood cells for culture. The specific megakaryocyte progenitor cell medium is a serum-free medium containing 40vol%-50vol% of hematopoietic stem cell culture supernatant, 50+/-10 ng/mL SCF and 50+/-10 ng/mL TPO, wherein the hematopoietic stem cell culture supernatant is obtained by culturing umbilical cord blood hematopoietic stem cells in a serum-free medium containing IL-6, IL-3, SCF, Flt-3L and TPO for 3-4 days. With the adoption of the method, the progenitor cell characteristics of umbilical cord blood megakaryocyte progenitor cells realizing in-vitro amplification are effectively improved, the megakaryocyte progenitor cell obtaining efficiency is improved, and accordingly, the capacity of the megakaryocyte progenitor cells differentiating into blood platelet is improved. In the amplification stage of the megakaryocyte progenitor cells, the consumption of added cell factors is reduced, accordingly, material consumption is reduced, and the cost is reduced.

The invention relates to the field of cell culture, in particular to a cell culture solution and application thereof to culture of megakaryocyte progenitor cells. According to the invention, WNT3a protein, insulin, Flt-3L, albumin, non-essential amino acid, thrombopoietin, stem cell growth factors, interleukin 3 and interleukin 6 are compounded, and a basal culture medium is added, so that richer nutrition can be provided for cells. Experiments show that the culture solution provided by the invention is used for culture of megakaryocyte progenitor cells and can selectively promote growth and proliferation of the megakaryocyte progenitor cells. After the culture solution provided by the invention cultures the megakaryocyte progenitor cells for 20 days, the number of cells is increased by 400-450 times, and the CD34+ and CD41+ double-positive expression rate in the cells is not lower than 35%. The effect is remarkably superior to that of a culture medium in the prior art (p is less than 0.01).

Isolated thrombopoietin (TPO), isolated DNA encoding TPO, and recombinant or synthetic methods of preparing and purifying TPO are disclosed. Various forms of TPO are shown to influence the replication, differentiation or maturation of blood cells, especially megakaryocytes and megakaryocyte progenitor cells. Accordingly, these compounds may be used for treatment of thrombocytopenia.

Isolated mpl ligand, isolated DNA encoding mpl ligand, and recombinant or synthetic methods of preparing mpl ligand are disclosed. These mpl ligands are shown to influence the replication, differentiation or maturation of blood cells, especially megakaryocytes and megakaryocyte progenitor cells. Accordingly, these compounds may be used for treatment of thrombocytopenia.