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Method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells

A technique for islet-like cells and stromal cells, which is applied in the field of inducing the differentiation of placental mesenchymal stem cells into islet-like cells, can solve the problems of reducing the number of bone marrow stem cells, difficulty in obtaining materials, and easily causing tumors, and achieves a wide range of placenta sources and expansion ability. Strong effect with less chance of infection

Inactive Publication Date: 2015-12-09
SHANDONG NEW MEDICINE RES INST OF INTEGRATED WESTERN MEDICINE & TCM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Theoretically speaking, embryonic stem cells (ESCs) are a group of primitive cells with the characteristics of totipotent stem cells, but they are prone to cause tumors, and there are ethical disputes; bone marrow mesenchymal stem cells are also a hot spot in stem cell research , is currently the most widely used stem cell, but the number of bone marrow stem cells will gradually decrease due to the aging of the individual, and the differentiation function will also decrease with age, and it is difficult to obtain materials, so it is urgent to find a new source of stem cells
[0003] The content of mesenchymal stem cells in bone marrow is very low, generally 0.001%-0.01%, and direct analysis of materials cannot meet the requirements; embryonic stem cells (ESCs) are a group of relatively primitive cells with the characteristics of totipotent stem cells, but they are easy to cause tumors Occurrence, and there is controversy over ethical issues

Method used

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  • Method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells
  • Method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells
  • Method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells

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Effect test

Embodiment 1

[0036] The method for inducing placental mesenchymal stem cells to differentiate into islet-like cells, the steps are as follows:

[0037] Step 1: The stage of in vitro isolation and culture of placental mesenchymal stem cells, as follows:

[0038] Select the placenta of full-term cesarean section, digest the placental tissue with 0.1% type Ⅰ collagenase for 60 minutes, centrifuge at 1800r / min for 5 minutes, obtain the digested cell pellet, wash the cell pellet twice, and inoculate it in ordinary culture In the medium, the medium contains 10% fetal bovine serum and α-MEM, and the culture conditions are 37°C, 5% CO2, and 95% saturated humidity. The medium is changed after 72 hours, and the medium is changed every 2 days. Once, the growth of the cells was observed under a microscope; the observations were as follows: figure 1 shown.

[0039] Step 2: Phenotyping stage of placental mesenchymal stem cells, as follows:

[0040] The vigorously growing cells of the third generation...

Embodiment 2

[0054] The method for inducing placental mesenchymal stem cells to differentiate into islet-like cells, the steps are as follows:

[0055] Step 1: The stage of in vitro isolation and culture of placental mesenchymal stem cells, as follows:

[0056] Select the placenta of full-term cesarean section, digest the placental tissue with 0.1% type Ⅰ collagenase for 60 minutes, centrifuge at 1800r / min for 5 minutes, obtain the digested cell pellet, wash the cell pellet twice, and inoculate it in ordinary culture In the medium, the medium contains 10% fetal bovine serum and α-MEM, and the culture conditions are 37°C, 5% CO2, and 95% saturated humidity. The medium is changed after 72 hours, and the medium is changed every 2.5 days. Once, the growth of the cells was observed under a microscope; the observations were as follows: figure 1 shown.

[0057] Step 2: Phenotyping stage of placental mesenchymal stem cells, as follows:

[0058] The vigorously growing cells of the third generati...

Embodiment 3

[0072] The method for inducing placental mesenchymal stem cells to differentiate into islet-like cells, the steps are as follows:

[0073] Step 1: The stage of in vitro isolation and culture of placental mesenchymal stem cells, as follows:

[0074] Select the placenta of full-term cesarean section, digest the placental tissue with 0.1% type Ⅰ collagenase for 60 minutes, centrifuge at 1800r / min for 5 minutes, obtain the digested cell pellet, wash the cell pellet three times, and inoculate it in ordinary culture In the medium, the medium contains 10% fetal bovine serum and α-MEM, and the culture conditions are 37°C, 5% CO2, and 95% saturated humidity. The medium is changed after 72 hours, and the medium is changed every 3 days. Once, the growth of the cells was observed under a microscope; the observations were as follows: figure 1 shown.

[0075] Step 2: Phenotyping stage of placental mesenchymal stem cells, as follows:

[0076] The vigorously growing cells of the third gene...

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Abstract

The invention relates to a method for inducing placenta-derived mesenchymal stem cells to be differentiated into islet-like cells. According to the method, placenta-derived mesenchymal stem cells are induced to be differentiated to islet-like cells and are identified by using cytokines, such as basic fibroblast growth factors (bFGF), hexadecadrol and beta-mercaptoethanol. The method lies in further investigating the differentiation potential of placenta-derived mesenchymal stem cells, discussing and studying optimum conditions of in-vitro separation, culture and differentiation of the placenta-derived mesenchymal stem cells, studying the possibility and the feasibility of inducing the placenta-derived mesenchymal stem cells to be differentiated into the islet-like cells, seeking an optimum inducing system of inducing differentiation in vitro and further finding a method for inducing the placenta-derived mesenchymal stem cells to be differentiated to insulin-producing cells (IPCs). Therefore, the method provides a foundational research basis for clinical application of the placenta-derived mesenchymal stem cells to the insulin-producing cells.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for inducing placental mesenchymal stem cells to differentiate into islet-like cells. Background technique [0002] In recent years, many scholars are actively looking for sources of stem cells that can differentiate into insulin-secreting cells. Stem cells currently used in research on islet cell sources mainly include embryonic stem cells and adult stem cells. Theoretically speaking, embryonic stem cells (ESCs) are a group of primitive cells with the characteristics of totipotent stem cells, but they are prone to cause tumors, and there are ethical disputes; bone marrow mesenchymal stem cells are also a hot spot in stem cell research , is currently the most widely used stem cell, but the number of bone marrow stem cells will gradually decrease due to the aging of the individual, and the differentiation function will also decrease with age, and it is di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0735C12N5/0775
Inventor 王泰华
Owner SHANDONG NEW MEDICINE RES INST OF INTEGRATED WESTERN MEDICINE & TCM CO LTD
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