Method for directionally inducing insulin-secreting cells by endometrial stem cells

A technology of endometrial stem cells and insulin secretion, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problem that stem cells can not be ignored when transforming into tumor cells

Inactive Publication Date: 2011-08-17
项桂法
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Problems solved by technology

[0023] At present, there is only one research literature that has reported the study of pancreatic secretory cells induced by endometrial stem cells, but the cells used in this liter

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  • Method for directionally inducing insulin-secreting cells by endometrial stem cells
  • Method for directionally inducing insulin-secreting cells by endometrial stem cells
  • Method for directionally inducing insulin-secreting cells by endometrial stem cells

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Embodiment Construction

[0073] 1. Isolation, culture and expansion of endometrial stem cells:

[0074] Ten menstruating women of different ages were recruited to donate menstrual blood samples on a completely voluntary basis.

[0075] The specific operating procedures are as follows:

[0076] 1. Collection of menstrual blood samples:

[0077]Put the menstrual blood collection tube and menstrual cup into the menstrual blood collection box and send it to the menstrual blood donor. The collection tube was pre-filled with 20ml of HBSS (Hank's Balanced Salt Solution) collection solution, which contained Vancomycin (vancomycin) 80 μg / mL, Claforan (cephalexin) 250 μg / mL, Amikacin (kanamycin) 100 μg / mL, Gentamycin (Gentamicin) 120 μg / mL, Amphotericin B (amphotericin B) 2.7 μg / mL and 400 units of heparin sodium. Use this as a sterilizing solution containing antibiotics.

[0078] That is, the preparation method of the above-mentioned HBSS (Hank's balanced salt solution) collection solution is as follows: ...

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Abstract

The invention discloses a method for directionally inducing insulin-secreting cells by endometrial stem cells, which comprises the following steps of: 1, separation culture and amplification of the endometrial stem cells; and 2, in vitro inducing differentiation. The step of in vitro inducing differentiation comprises the following specific steps of: digesting the fourth to sixth generation of cells with good growth conditions, which are obtained in the step 1, by pancreatic enzymes; when the cells are observed under a microscope to be changed into single circles, stopping digesting by a DMEM(dulbecco's modified eagle medium) culture medium containing fetal calf serum with the volume concentration of 13 to 17 percent; centrifuging and washing the obtained cell precipitates by a PBS (Phosphate Buffer Solution); adding an inducing culture medium into the cells washed by the PBS; placing the obtained cells into an incubator with the concentration of carbon dioxide CO2 of 4 to 6 percent,the saturated humidity of 94 to 96 percent and the temperature of 36 to 38 DEG C for culturing; and culturing the cells for 13 to 15 days to obtain the insulin-secreting cells. The obtained insulin-secreting cells are used for treating diabetes mellitus so as to provide a novel seed resource for stem cell therapy.

Description

technical field [0001] The invention relates to a method for directional induction of insulin-secreting cells by endometrial stem cells. Background technique [0002] Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia caused by defects in insulin secretion or action, or both. Clinically, it is divided into type 1, type 2, other special types, and gestational diabetes. Type 1 diabetes is an insulin-dependent diabetes mellitus, an autoimmune disease mainly due to the destruction of the cells characteristic of insulin. During the onset stage, the β insulin-secreting cells in the islets of the pancreas are selectively destroyed by autoimmune attacks, unable to produce enough insulin, and the regulation of blood sugar concentration is out of balance, resulting in hyperglycemia. According to the survey results of the 19th International Diabetes Congress in 2006, the average incidence of diabetes in the world is about 5.7% or more, and there are abou...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
Inventor 项春生
Owner 项桂法
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