1. A manufacturing method of a semiconductor device, comprising:
(a) setting up a paste including a resin on an electrical coupling part coupled electrically to a semiconductor substrate;
(b) setting up a soldering material over the electrical coupling part in contact with the paste; and
(c) forming an external terminal from the soldering material and forming a reinforcement from the paste by fusing the soldering material and the paste, wherein the reinforcement is formed such that part of the external terminal is exposed and a periphery of an edge of a base coupled to the electrical connection part of the external terminal is covered.
2. The manufacturing method of the semiconductor device according to claim 1, wherein the paste is set up in a manner of covering the entire electrical connection part in the step (a).
3. The manufacturing method of the semiconductor device according to claim 1, wherein the paste is set up respectively on a plurality of the electrical coupling parts such that one paste is not in contact with another adjacent paste in the step (a).
4. The manufacturing method of the semiconductor device according to claim 1, wherein the paste is set up integrally over the plurality of the electrical coupling parts in the step (a).
5. The manufacturing method of the semiconductor device according to claim 1, wherein the soldering material is set up such as to be not in contact with the electrical connection part in the step (b).
6. A manufacturing method of a semiconductor device, comprising:
(a) depositing a paste including a resin on a soldering, material;
(b) setting up the soldering material such that the paste is in contact with the electrical connection part which is electrically coupled to the semiconductor substrate; and
(c) forming an external terminal from the soldering material by fusing the soldering material and the paste and forming the reinforcement from the paste, wherein the reinforcement is formed such that part of the external terminal is exposed and a periphery of an edge of a base coupled to the electrical connection part of the external terminal is covered.
7. The manufacturing method of the semiconductor device according to claim 6, wherein the electrical connection part is part of an interposer mounted on the semiconductor substrate.
8. The manufacturing method of the semiconductor device according to claim 6, wherein the electrical connection part is part of a wiring layer extending over a resin layer formed on the semiconductor substrate.
9. The manufacturing method of the semiconductor device according to claim 6, wherein the paste includes the flux.
10. The manufacturing method of the semiconductor device according to claim 6, wherein the paste includes the soldering material.
11. The manufacturing method of the semiconductor device according to claim 1, wherein the electrical connection part is part of an interposer mounted on the semiconductor substrate.
12. The manufacturing method of the semiconductor device according to claim 1, wherein the electrical connection part is part of a wiring layer extending over a resin layer formed on the semiconductor substrate.
13. The manufacturing method of the semiconductor device according to claim 1, wherein the paste includes the flux.
14. The manufacturing method of the semiconductor device according to claim 1, wherein the paste includes the soldering material.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method of culturing human embryonic stem cells into erythroid cells producing adult-type \u03b2-hemoglobin, the method comprising the steps of:
(a) isolating human embryonic stem cell-derived cells selected from the group consisting of CD31+, CD34+, and CD31+CD34+ cells, wherein the cells are enriched in CD31+CD43+, CD31+CD34+, CD34+CD43+ or CD31+CD34+CD43+ hematopoietic progenitors;
(b) culturing the hematopoietic progenitors to cause an expansion of erythroid precursors; and
(c) recovering a population of cells, wherein essentially all of the live cells are erythroid cells that produce adult \u03b2-hemoglobin but not embryonic \u03b6-hemoglobin.
2. The method of claim 1, wherein the cells of step (c) are essentially free of lymphocytes.
3. The method of claim 1, wherein the cells of step (c) contain less than 0.1% leukocytes.
4. The method of claim 1, wherein the culturing of step (b) is in the presence of stem cell factor and erythropoietin under conditions preventing cell adherence.
5. The method of claim 1, wherein the human embryonic stem cells are co-cultured with stromal cells to produce the hematopoietic progenitors.
6. The method of claim 1, wherein at least 0.2\xd7105 erythroid cells are generated from one human embryonic stem cell.
7. The method of claim 1, wherein up to 2.0\xd7105 erythroid cells are generated from one human embryonic stem cell.
8. The method of claim 1, wherein the cells of step (c) produce adult-type \u03b2-hemoglobin, are CD31\u2212, CD34\u2212, CD71+, CD235a+ and additionally express fetal \u03b3-hemoglobin.
9. The method of claim 8, wherein the cells of step (c) do not express embryonic \u03b6-hemoglobin.
10. The method of claim 1, wherein the purity of the isolated hematopoietic progenitors are greater than 95% at a single column run and cell viability, as evaluated by Trypan blue exclusion, is higher than 95%.
11. The method of claim 1, wherein the CD31+CD34+ cells comprise 30% to 50% CD31+CD34+CD43+ hematopoietic progenitors and 50% to 70% CD31+CD34+CD43\u2212KDRbright endothelial cells.
12. The method of claim 1, wherein the cells express adult \u03b2-hemoglobin by 10 days (\xb110%) of culture, and wherein most of the cells have a phenotype and a morphology of erythroid cells.
13. The method of claim 1, wherein after approximately 30 days (\xb110%) of culture, essentially all of the live cells (>95%) show positive stains with antibodies against fetal \u03b3-hemoglobin and adult \u03b2-hemoglobin, but no positive staining with antibodies against embryonic \u03b6-hemoglobin.
14. The method of claim 1, wherein after approximately 50 days (\xb110%) of culture, the cells of step (c) are CD31\u2212CD34\u2212 and show adult \u03b2-hemoglobin expression and no embryonic \u03b6-hemoglobin expression.
15. The method of claim 1, wherein the hematopoietic precursors are cultured in serum-free culture conditions.
16. A cell population of step (c) of claim 1.
17. The cell population of claim 16, wherein essentially all live cells are nucleated erythroid cells.
18. The cell population of claim 16, wherein the population is essentially free of lymphocytes.
19. The cell population of claim 16, wherein the population contains less than 0.1% leukocytes.
20. The cell population of claim 16, wherein the population is CD31\u2212, CD34\u2212, CD71+ and CD235a+.
21. A preparation of CD31+CD34+CD43+ hematopoietic progenitor cells.
22. The preparation of claim 21, wherein the cells are the result of 6 to 7 days (\xb110%) co-culture between human embryonic stem cells and OP9 cells.
23. A CD31+ cell population, comprising 10% to 20% CD31+CD43+ hematopoietic progenitors, up to 60% CD31+CD43\u2212KDRbright endothelial cells and less than 15% CD34+CD43\u2212KDR-mesenchymal cells.
24. A CD34+ cell population, comprising 10% to 20% CD34+CD43+ hematopoietic progenitors, up to 60% CD34+CD43\u2192KDRbright endothelial cells and less than 15% CD34+CD43\u2212KDR-mesenchymal cells.