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In-vitro efficient amplification agent for adipose tissue-derived stromal cells and amplification method

A technology of mesenchymal stem cells and in vitro expansion, applied in the biological field, can solve problems such as inability to realize industrialization and difficulty in meeting clinical needs

Active Publication Date: 2019-05-31
上海揽微赛尔生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on the legality, clinical treatment of ADSCs for plastic surgery in Korea and Japan is not restricted, but all the studies conducted are mostly in the early stages
[0004] Although the preparation of ADSCs is relatively simple, and the in vitro proliferation rate is faster than that of other sources of mesenchymal stem cells, but because a large number of ADSCs are often required for clinical large-scale surgery, it is difficult to meet clinical needs by relying on natural proliferation, and it is impossible to achieve industrialization.

Method used

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  • In-vitro efficient amplification agent for adipose tissue-derived stromal cells and amplification method
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  • In-vitro efficient amplification agent for adipose tissue-derived stromal cells and amplification method

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Embodiment Construction

[0019] The following describes the substantive content of the present invention in detail, but does not limit the protection scope of the present invention.

[0020] 1. Experimental materials

[0021] Clean-grade domestic rabbits, male or female, weighing about 1.2kg, will be used after one week of adaptive feeding.

[0022] Fetal bovine serum and DMEM medium were produced by Gibo; CD14 was produced by CALTAG; CD29 and CD44 were produced by CHEMCON; CD45 was produced by ANTIGENIX.

[0023] Glycoursodeoxycholic acid HPLC purity ≥ 95%.

[0024] 2. Experimental method

[0025] 1. Isolation, preparation, culture and phenotypic identification of ADSCs

[0026] After anesthetizing the rabbit, take out the adipose tissue in the inguinal groove on both sides, cut it into pieces and homogenate, digest with 0.075% collagenase Ⅰ, pass through a 300-mesh sieve, centrifuge at 1200r / min for 5min at 4°C, and discard the upper layer Fat and supernatant, the precipitate was first washed 3 ...

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Abstract

The invention discloses an in-vitro efficient amplification agent for adipose tissue-derived stromal cells and an amplification method. It is found that glycoursodeoxycholic acid has a remarkable proliferation promoting effect on the ADSCs and has a certain dosage dependent effect, and the glycoursodeoxycholic acid cannot reduce the multi-directional differentiation potential of the ADSCs. Therefore, the glycoursodeoxycholic acid can be used as the in-vitro efficient amplification agent for the adipose tissue-derived stromal cells, used for preparing an in-vitro amplification culture medium ofthe adipose tissue-derived stromal cells and used for in-vitro amplification of the adipose tissue-derived stromal cells.

Description

technical field [0001] The invention belongs to the biological field, and relates to in vitro culture and expansion of adipose-derived mesenchymal stem cells, in particular to an in vitro high-efficiency expansion agent and an expansion method for adipose-derived mesenchymal stem cells. Background technique [0002] In the past, fat was only regarded as an energy storage tissue and did not attract the attention of researchers. Until the mid-1980s, after it was determined that it was involved in the metabolism of sex hormones, adipose tissue became an important endocrine organ involved in human metabolism and immune response. In 2001, Zuk et al first reported a new source of adult stem cells, adipose-derived stem cells (ADSCs) isolated from adipose tissue, and reached a consensus in 2004 called ADSCs. Since then, because of its high proliferation and self-renewal ability like bone marrow-derived stem cells (BMSCs), a large number of ADSCs can be isolated from adipose tissue,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 不公告发明人
Owner 上海揽微赛尔生物科技有限公司
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