Method and system for introducing a gene into a human stem cell
a technology of stem cell and gene, applied in the field of vector systems for gene and stem cell therapy, can solve the problems of multiple organ failure and death, increased risk of infection, and increased risk of infection, and achieve the effect of improving the survival rate of patients, improving the survival rate, and improving the survival ra
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example i
[0029] Human oocytes may be collected from an adult female that preferably has given appropriate informed consent to the use of the oocytes. Techniques for manipulation of human oocytes are well known in the art of infertility treatment. Briefly, ovarian hyperstimulation may be initiated with leuprolide acetate (Lupron; TAP Pharmaceuticals, North Chicago, Ill.) and administered from the midluteal phase at 1.0 mg / day subcutaneously for 10 days. The dosage may then be reduced to 0.5 mg / day until the morning of human chorionic gonadotropin (hCG) injection. After pituitary down-regulation of estrogen, human menopausal gonadotropin may be given until at least two follicles have reached a mean diameter of 1.8 cm. Oocyte retrieval may be scheduled for 35 hours after the hCG injection.
[0030] Human sperm cells may be collected from an adult male that preferably has also given appropriate informed consent to the use of his sperm cells. The ejaculated sperm cells may be collected and prepared...
example ii
[0031] This example illustrates the ability of the linker antibody, mAbC, to bind to human sperm cells. Briefly, human sperm cells were incubated with the different primary monoclonal antibodies, washed, and further incubated with a secondary antibody that specifically recognized mouse immunoglobulin. This secondary antibody, which was commercially available and well known in the art, had fluorescent molecules such as fluorescein or rhodamine conjugated to it. Once the secondary antibody molecules were bound and washed, a flow-cytometry instrument or a FACS sorter counted the number of fluorescent sperm cells with bound primary and secondary antibodies from naked sperm cells.
[0032]FIGS. 2-4 show flow-cytometry analyses for mAbC, linker antibody that binds to the sperm cells from human, mouse, and pig, respectively. mAbC also binds to the sperm cells of cow, chicken, goat, and sheep, which are disclosed in U.S. patent application Ser. No. 09 / 537,861. The Y-axis of the figures corres...
example iii
[0036] This example illustrates the in vitro fertilization (IVF) procedure and the culturing of the resulting zygotes into embryonic stem cells. Human sperm cells may be incubated with the mAbC linker and any DNA molecule using similar techniques as described in mouse and pigs. IVF of the isolated human oocytes may then be performed with the human sperm-linker-DNA complexes using procedures routinely used in fertility clinics.
[0037] Resulting zygotes may then be cultured into human blastocysts using G1 media from day 0 to day 3 and switching to G2 media from day 3 to 5, as reported by Gardner, supra. Table 2 provides the composition of Gardner's G1 and G2 media based on Barnes, F. L., et. al. (1995) “Blastocyst development and pregnancy after in vitro maturation of human primary oocytes, intracytoplasmic sperm injection and assisted hatching,”Human Reproduction, 10:3243-3247, which is hereby incorporated by reference in its entirety as if fully set forth herein.
[0038] Concentratio...
PUM
| Property | Measurement | Unit |
|---|---|---|
| diameter | aaaaa | aaaaa |
| immunological compatibility | aaaaa | aaaaa |
| immunologically compatible | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More