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A kind of preparation method of human placenta collagen

A human placenta and collagen technology, applied in the field of biomedical material preparation, can solve the problems of difficult control of key links of product quality, unstable product quality, affecting the application effect and value of collagen and collagen-based biomedical materials, etc.

Active Publication Date: 2016-08-10
SICHUAN UNIV
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Problems solved by technology

[0008] 2. The existing collagen extraction technology has a long cycle, requiring more than 15 days, and the steps are cumbersome, so that the extraction efficiency and yield are relatively low, and the key links in the process that affect product quality are difficult to control, which may easily lead to unstable product quality and ultimately affect collagen. And the application effect and value of collagen-based biomedical materials

Method used

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  • A kind of preparation method of human placenta collagen
  • A kind of preparation method of human placenta collagen

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preparation example Construction

[0046] The preparation process of the present invention is as follows: human placenta tissue is used as raw material, followed by pretreatment, enzyme digestion and graded salting out.

[0047] The pretreatment steps include fully cleaning the placenta tissue and cutting it into small pieces, centrifuging to remove the supernatant after homogenizing at low temperature, repeatedly washing and centrifuging with deionized water until the supernatant is transparent; the precipitation contains metal complexing agents and neutral salts The pretreatment buffer was soaked and stirred slowly and then centrifuged, the obtained precipitate was washed once with the pretreatment buffer and centrifuged, and the precipitate was taken for later use; the enzyme digestion step included soaking the pretreated human placenta tissue in the digestive solution, and the low temperature condition Under continuous slow stirring, after enzymatic digestion for 6-30h, centrifuge and take the supernatant fo...

Embodiment 1

[0050] Extract collagen as follows:

[0051] (1) Use fresh intact human placenta obtained through disease screening and normal delivery;

[0052] (2) Fully wash the placenta tissue and cut it into small pieces, centrifuge to remove the supernatant after homogenizing at low temperature, wash the precipitate with deionized water and then centrifuge, repeat the cleaning and centrifugation for 4 times until the supernatant is transparent; the pH for precipitation is 7.2- 7.4. Soak in 0.5 mol / L tris-hydrochloric acid (Tris-HCl) buffer solution containing 10 mmol / L sodium ethylenediaminetetraacetic acid (Na2EDTA) and 1.0 mol / L NaCl and stir slowly for 4 hours, then stir The speed should not be too fast, it is advisable to rotate the liquid surface slightly; after the stirring is completed, centrifuge, and the precipitate obtained after centrifugation is washed once with Tris-HCl buffer solution and centrifuged, and finally the precipitate obtained by centrifugation is weighed and us...

Embodiment 2

[0072] Extract collagen as follows:

[0073] (1) Use fresh intact human placenta obtained through disease screening and normal delivery;

[0074] (2) The placental tissue was fully cleaned and cut into small pieces, and the supernatant was removed by centrifugation after homogenization at low temperature. The precipitate was washed with deionized water and then centrifuged. Repeated cleaning and centrifugation 4 times until the supernatant was transparent; the pH of the precipitation was 7.2, Contains 20 mmol / L sodium edetate (Na 2 EDTA), 2.0 mol / L NaCl in 0.05 mol / L Tris-HCl buffer solution and stir slowly for 4 hours. Centrifuge after the stirring is completed. After centrifugation, the precipitate obtained is washed once with Tris-HCl buffer and centrifuged. Finally, the precipitate obtained by centrifugation is weighed and used as the basis for subsequent measurement;

[0075] (3) Soak the precipitate finally obtained in step (2) in the digestive solution, which is 0.8mo...

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Abstract

The present invention relates to a preparation method of human placenta collagen. The method uses human placenta tissue as raw material, and sequentially performs pretreatment, enzyme digestion, and graded salting-out to obtain human type I, type III, type IV and At least one of the V-type medical collagen materials. The method greatly shortens the preparation cycle of human placenta type I, type III, type IV and type V collagen materials, improves the preparation efficiency and yield, and makes the quality of the product more stable at the same time. The method can also be used according to actual needs. By extracting specific types of collagen, human type I, type III, type IV and type V collagen materials with natural structures can also be obtained at the same time, making full use of human placental tissue resources.

Description

technical field [0001] The invention relates to a method for preparing human placenta type I, type III, type IV and type V collagen materials maintaining natural structure by using healthy human placenta obtained after normal delivery as a raw material, belonging to the field of biomedical material preparation. Background technique [0002] As a specific structural protein, natural collagen has been widely recognized for its characteristics and advantages, and has further become one of the important raw materials for biomedical materials. At least 26 collagen types are currently known in the human body, each of which adds a specific function to collagen's structural role. Mammalian placenta, especially human placenta, is very rich in collagen, including type I, type III, type IV and type V collagen, so it is a very valuable raw material for preparing natural medical collagen. [0003] Various types of natural medical collagen prepared using human placenta as raw materials d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/06C07K14/78C07K1/30
Inventor 林海樊渝江张兴栋
Owner SICHUAN UNIV
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