Methods of inducing pluripotent adipose-derived stem cells

A fat stem cell, multipotential technology, applied in the field of bioengineering, can solve the problems of long time-consuming, expensive, and small quantity, and achieve the effect of promoting fracture healing, long treatment cycle, and many complications

Active Publication Date: 2020-03-06
河南创新生物科技研究院有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional method of extracting bone marrow will bring some pain to patients, usually requires general anesthesia or spinal anesthesia, and the quantity is small, and the expansion in vitro takes a long time and is expensive.

Method used

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  • Methods of inducing pluripotent adipose-derived stem cells
  • Methods of inducing pluripotent adipose-derived stem cells
  • Methods of inducing pluripotent adipose-derived stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042]A method for inducing pluripotent fat stem cells, comprising:

[0043] (1) Isolation and culture of rat ADSCs:

[0044] Reference (Zuk P A, Zhu M, Mizuno H, et al. Multilineage cells from humanadipose tissue: implications for cell-based therapies[J]. TissueEng, 2001, 7:211-28.) method: take 1 healthy 3 - 4-week-old SD rats were anesthetized with 10% chloral hydrate and soaked in 75% alcohol for 15 minutes. The inguinal subcutaneous fat of the rats was separated on a clean bench, and washed 3 times with PBS containing 1% double antibody. After the adipose tissue was cut into small pieces, double volume of 0.1% type Ⅰ collagenase solution was added and mixed evenly. Place the sealed centrifuge tube in a constant temperature water bath at 37°C for 60 minutes with slow shaking and digestion, then centrifuge at 1200r / min for 20 minutes to remove the upper layer of fat, undigested adipose tissue and fibrous connective tissue. Cells were resuspended in phosphate buffer saline...

Embodiment 2

[0051] OricellTMSD Rat Adipose-derived Mesenchymal Stem Cell Osteogenic Induction and Differentiation Medium Kit (Cyagen, China, Cat. No.: RAMSD-90021) was used as a control.

[0052] A method for inducing pluripotent fat stem cells, comprising:

[0053] (1) Isolation and culture of rat ADSCs:

[0054] Reference (Zuk P A, Zhu M, Mizuno H, et al. Multilineage cells from humanadipose tissue: implications for cell-based therapies[J]. TissueEng, 2001, 7:211-28.) method: take 1 healthy 3 - 4-week-old SD rats were anesthetized with 10% chloral hydrate and soaked in 75% alcohol for 15 minutes. The inguinal subcutaneous fat of the rats was separated on a clean bench, and washed 3 times with PBS containing 1% double antibody. After the adipose tissue was cut into small pieces, double volume of 0.1% type Ⅰ collagenase solution was added and mixed evenly. Place the sealed centrifuge tube in a constant temperature water bath at 37°C for 60 minutes with slow shaking and digestion, then c...

Embodiment 3

[0060] DMEM medium was used as a negative control.

[0061] A method for inducing pluripotent fat stem cells, comprising:

[0062] (1) Isolation and culture of rat ADSCs:

[0063] Reference (Zuk P A, Zhu M, Mizuno H, et al. Multilineage cells from humanadipose tissue: implications for cell-based therapies[J]. TissueEng, 2001, 7:211-28.) method: take 1 healthy 3 - 4-week-old SD rats were anesthetized with 10% chloral hydrate and soaked in 75% alcohol for 15 minutes. The inguinal subcutaneous fat of the rats was separated on a clean bench, and washed 3 times with PBS containing 1% double antibody. After the adipose tissue was cut into small pieces, double volume of 0.1% type Ⅰ collagenase solution was added and mixed evenly. Place the sealed centrifuge tube in a constant temperature water bath at 37°C for 60 minutes with slow shaking and digestion, then centrifuge at 1200r / min for 20 minutes to remove the upper layer of fat, undigested adipose tissue and fibrous connective tis...

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Abstract

The invention discloses a method for inducing pluripotent adipose-derived stem cells. Ganoderma resinaceum element F is applied in the culture process. The method comprises the following steps: (1) carrying out isolated culture on rat ADSCs; and (2) osteogenesis induced differentiation: taking third-generation ADSCs cells with a good growth state, performing conventional digestion, terminating digestion, centrifuging and cell resuspending, inoculating the obtained cells into a six-pore plate with preset cover glass in a form of 100/pore, changing an experimental group into an osteogenesis induction culture medium 1 for osteogenesis induction culture for 1 day after the cells are completely attached to the walls, changing a liquid on the second day, changing into an osteogenesis induction culture medium 2, and continuing osteogenesis induction for 7 days. According to the invention, the ganoderma resinaceum element F is innovatively applied to inducing pluripotent adipose-derived stem cells to be rapidly and efficiently differentiated into osteoblasts (8d), and since the ganoderma resinaceum element F is derived from ganoderma resinaceum, the long-term safety of the stemless ganoderma resinaceum element F is guaranteed, and the ganoderma resinaceum element F has the potential of being directly applied in vivo.

Description

technical field [0001] The invention relates to a method for inducing pluripotent fat stem cells, belonging to the technical field of bioengineering. Background technique [0002] Adult stem cells refer to undifferentiated cells existing in a differentiated tissue, which can self-renew and differentiate to form cells of the tissue. Adult stem cells exist in various tissues and organs of the body. Adult stem cells in adult individual tissues are mostly in a dormant state under normal conditions, and can exhibit different degrees of regeneration and renewal capabilities in pathological conditions or induced by external factors. Compared with embryonic stem cells, adult stem cells have limited proliferation ability. In terms of differentiation potential, embryonic stem cells are pluripotent and can form a variety of cells in the body, while adult stem cells can only differentiate into cell types of certain specific tissues. Adult stem cells are ubiquitous, the problem is how...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0654C12N2500/38C12N2501/39C12N2501/999C12N2506/1384
Inventor 韦丹李晗吴雷振秦志华任翔潘肖芬
Owner 河南创新生物科技研究院有限公司
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