Method for improving efficiency of mammalian somatic cell cloning technology
A technology of mammals and mammals, applied in botany equipment and methods, biochemical equipment and methods, plant genetic improvement, etc., can solve the problems of inconvenient industrial production, large number of requirements, cumbersome operation steps, etc., to reduce work intensity , improve efficiency, and simplify the effect of the cultivation process
Inactive Publication Date: 2015-11-18
AGRI GENOME INST OF SHENZHEN CHINESE ACADEMY OF AGRI SCI
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Problems solved by technology
However, the existing porcine somatic cell cloning technology is difficult to meet the requirements of large-scale industrial production. Large-scale preparation of cloned pigs requires a large number of cloned embryos. However, the operation efficiency of embryos is low, the required quantity is large, and the operation steps are cumbersome, which is not convenient for industrial production.
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Embodiment 1
[0030] Embodiment 1 A kind of method for improving the technical efficiency of porcine somatic cell cloning
[0031] A method for improving the efficiency of porcine somatic cell cloning technology of the present embodiment comprises the following steps:
[0032] 1. In vitro maturation culture of porcine oocytes
[0033] ①. After collecting the ovaries from the slaughterhouse, store them in 34-37°C normal saline containing 200 U / ml penicillin (Sigma, P3032) and 200 U / ml streptomycin (Sigma, S6501), and transport them within 2-3 hours to laboratory.
[0034] ②. Wash 3-4 times with normal saline containing penicillin (Sigma, P3032) 200U / ml and streptomycin (Sigma, S6501) 200U / ml incubated at 38°C to wash away blood and other pollutants.
[0035] ③.Use a 10ml disposable syringe with a 18-gauge needle, with the needle port facing downward, to collect the oocyte complex and follicular fluid in the follicles of 3-6mm from the side tissue.
[0036] ④. Collect the collected follicl...
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The invention discloses a method for improving the efficiency of mammalian somatic cell cloning. According to the method, oocytes are denucleate, then a hole is punched at any position of a denucleated oocyte transparent tape to carry out nuclear transplantation. The method separates denucleation operation from nuclear transplantation, so that mature oocyte picking, denucleation, nuclear transplantation and fusion-activation can be carried out on one pipeline at the same time, so that the somatic cell cloning technology can be industrialized, the operation can form a pipelining operation mode, and each operator only completes one operation action. Under the circumstance of the same personnel, the cloned reconstructed embryos prepared in one workday are 3 times that of the original technology. The microoperation efficiency is significantly enhanced, the work intensity of technicians is reduced, the training process of technicians is simplified, and the somatic cell cloning technology can be popularized.
Description
technical field [0001] The invention belongs to the field of biotechnology, and more specifically, the invention relates to a method for improving the efficiency of mammalian somatic cell cloning technology. Background technique [0002] The basic premise of animal somatic cell cloning is that a single cell has totipotency as a donor, which is transplanted into a mature oocyte from which the genetic material has been removed, and after activation, it develops into a cloned embryo in vitro and transplants it into a recipient animal, and produces the embryo after pregnancy Cloned animals with identical genetic information. The success of animal somatic cell cloning technology has played an important role in promoting the development of life sciences. It not only proves that animal somatic cells are totipotent, but also has great application prospects. With the continuous development in recent years, somatic cell cloning technology provides new means for studying the mechanism...
Claims
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Login to View More IPC IPC(8): C12N15/877A01K67/027
Inventor 李林李奎樊俊华唐中林白立景
Owner AGRI GENOME INST OF SHENZHEN CHINESE ACADEMY OF AGRI SCI