Method for inducing and differentiating functional cardiocytes by utilizing endometrial stem cells

An endometrial stem cell and cardiomyocyte technology, applied in animal cells, vertebrate cells, artificial cell constructs, etc., can solve the problems of unsatisfactory transplantation effect, limited cell number, cell death, etc., and improve the clinical treatment effect of stem cells. , Wide application, high in vitro differentiation rate

Active Publication Date: 2011-09-28
HANGZHOU S EVANS BIOSCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1. Insufficient potential to differentiate into functional cells such as cardiomyocytes and neuron cells;
[0005] 2. Most of the cells died after transplantation;
[0006] 3. The curative effect of transplantation is not satisfactory;
[0007] 4. Cell Number Limitation

Method used

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  • Method for inducing and differentiating functional cardiocytes by utilizing endometrial stem cells
  • Method for inducing and differentiating functional cardiocytes by utilizing endometrial stem cells
  • Method for inducing and differentiating functional cardiocytes by utilizing endometrial stem cells

Examples

Experimental program
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Embodiment Construction

[0055] 1. Isolation, culture and expansion of endometrial stem cells:

[0056] Ten menstruating women of different ages were recruited to donate menstrual blood samples on a completely voluntary basis.

[0057] The specific operating procedures are as follows:

[0058] 1. Collection of menstrual blood samples:

[0059] 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.

[0060] 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 inducing and differentiating functional cardiocytes by utilizing endometrial stem cells. The method comprises the following steps of: 1) performing separation culture on the endometrial stem cells and amplifying; and 2) inducing and differentiating in vitro, namely digesting 4th to 6th generation of cells in good growth condition, which is obtained in step 1) by using pancreatin, terminating the digestion by using a Dulbecco modified eagle medium (DMEM) containing 13 to 17 volume percent fetal calf serum when the cells are changed into single circular shapes under a microscope, centrifuging, washing an obtained cell precipitate by using phosphate buffer solution (PBS), adding an induction culture medium into the cells washed by the PBS, putting into an incubator which contains 4 to 6 percent of CO2 and has the saturated humidity of between 94 and 96 percent at the temperature of between 36 and 38 DEG C, and culturing to obtain the functional cardiocytes, wherein the culture time is 68 to 76 hours. The obtained functional cardiocytes can be used for treating myocardial infarction.

Description

technical field [0001] The invention relates to a method for using endometrium stem cells to induce differentiation of functional cardiomyocytes. Background technique [0002] At present, the incidence of congestive heart failure is increasing year by year. There are 15 million patients worldwide, with 465,000 new cases each year. It is the leading cause of hospitalization for people over 65 years old and the leading cause of cardiovascular death. Cell transplantation therapy using stem cells as a seed resource can promote angiogenesis and myocardial regeneration in the infarct area, thereby improving cardiac function after myocardial infarction, and has become a research hotspot in the cardiovascular field. However, embryonic cardiomyocytes available for transplantation have limited proliferative ability and inconvenient materials, and skeletal muscle myoblasts cannot form gap junctions with cardiomyocytes and can cause arrhythmia, which makes this treatment method stagnant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/071
Inventor 项春生
Owner HANGZHOU S EVANS BIOSCI LTD
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