Culture system of stem cells

A stem cell culture and pluripotent stem cell technology, applied in the field of stem cell culture system, can solve the problems of unstable cell state, cell aging, etc., achieve good cell adhesion and value-added effects, promote long-term growth, and ensure stability and activity.

Pending Publication Date: 2020-12-04
江苏育瑞康生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Stem cell medium and stem cell serum-free nutrient supplements are used together, and the effect of cell adhesion and value proliferation is better; but the existing stem cell culture system is easy to cause cells to be unstable and aging;

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A stem cell culture system, including a humanized matrix and a humanized medium that support the long-term culture of human pluripotent stem cells; the humanized medium contains sodium chloride precipitation components of human plasma, 100 U / m liter of penicillin, and 100 μg / mL Streptomycin, 10mM non-essential amino acid NEAA, 10mM mercaptoethanol; the preparation steps of humanized matrix are as follows:

[0021] S1. Cut the fresh placenta into pieces, put it into water, and stir it at 8°C for 6 hours, then sieve it, and rinse it with clean water. The water temperature is 8°C, and the water change time is 6 hours. Second time, after the second filtration, suspend the retained tissue with cold 1.5M sodium chloride buffer and stir at 8°C; the configuration process of sodium chloride buffer is as follows: 40 grams of sodium chloride and 15 mL of (Hydroxymethyl)aminomethane buffer solution, be dissolved in Mi liters of iQ water, adjust p hour to 7.0, and settle to 500 mil...

Embodiment 2

[0027] A stem cell culture system, including a humanized matrix and a humanized medium that support the long-term culture of human pluripotent stem cells; the humanized medium contains sodium chloride precipitation components of human plasma, 100 U / m liter of penicillin, and 100 μg / mL Streptomycin, 10mM non-essential amino acid NEAA, 10mM mercaptoethanol; the preparation steps of humanized matrix are as follows:

[0028] S1. Cut the fresh placenta into blocks, put it into water, and stir it at 10°C for 8 hours, then sieve it, and rinse it with clean water. The water temperature is 10°C, and the water change time is 8 hours. Second time, after the second filtration, suspend the retained tissue with cold 1.5M sodium chloride buffer and stir at 10°C; the configuration process of sodium chloride buffer is as follows: 40 grams of sodium chloride and 15 mL of (Hydroxymethyl)aminomethane buffer solution, be dissolved in Mi liters of iQ water, adjust p hour to 7.0, and settle to 500 ...

Embodiment 3

[0034] A stem cell culture system, including a humanized matrix and a humanized medium that support the long-term culture of human pluripotent stem cells; the humanized medium contains sodium chloride precipitation components of human plasma, 100 U / m liter of penicillin, and 100 μg / mL Streptomycin, 10mM non-essential amino acid NEAA, 10mM mercaptoethanol; the preparation steps of humanized matrix are as follows:

[0035]S1. Cut the fresh placenta into pieces, put it into water, and stir it at 12°C for 10 hours, then sieve it, and rinse it with clean water. Second time, after the second filtration, suspend the retained tissue with cold 1.5M sodium chloride buffer and stir at 12°C; the configuration process of sodium chloride buffer is as follows: 40 grams of sodium chloride and 15 mL (Hydroxymethyl)aminomethane buffer solution, be dissolved in Mi liters of iQ water, adjust p hour to 7.0, and settle to 500 milliliters; Tris(hydroxymethyl)aminomethane buffer solution is as follo...

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Abstract

The present invention provides a culture system of stem cells and particularly relates to the technical field of cells. The culture system of the stems cells comprises a humanized matrix and a humanized culture medium for supporting long-term culture of human pluripotent stem cells; A preparation method of the humanized matrix comprises the following steps: S1, cutting fresh placenta into blocks,cleaning the placenta blocks twice and conducting filtering; S2, conducting washing with a cold 1M sodium chloride buffer solution, and conducting washing with clean water for 6 times, wherein water is replaced with clean water for 6-10 hours once ; S3, conducting washing with a cold 0.3 M acetic acid solution for 3 days, whererin the solution is replaced once every 6-10 hours, and conducting filtering and weighing after the 0.3 M acetic acid solution is used for the ninth time; S4, centrifuging the weighed substance for 1 hour at a temperature of 8-12 DEG C, removing a supernatant, collectinga precipitate, conducting weighing, adding a 1.8 M urea buffer solution according to a mass-volume ratio of 1:1, and conducting stirring for 24 hours at a temperature of 8-12 DEG C; and S5, conducting centrifuging for 30 min at a temperature of 8-12 DEG C, adding the supernatant into a 30KD dialysis bag, and conducting dialyzing to obtain liquid, namely the humanized matrix. The culture system can ensure stable cell state and activity.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a stem cell culture system. Background technique [0002] Stem cells are a class of cells with unlimited or immortal self-renewal capacity, capable of producing at least one type of highly differentiated progeny cells. Over the years, the definition of stem cells has been continuously revised and defined from different levels. At present, most biologists and medical scientists believe that stem cells are a type of cells that come from embryos, fetuses or adults with unlimited self-renewal, proliferation and differentiation capabilities under certain conditions, and can produce phenotypes and genotypes that are exactly the same as themselves. Daughter cells can also produce specialized cells that make up body tissues and organs, and can also differentiate into progenitor cells. Stem cell culture promotes long-term growth of human stem cells while maintaining multilinea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074
CPCC12N5/0696C12N2533/90C12N2500/44C12N2500/90C12N2500/32
Inventor 王建波
Owner 江苏育瑞康生物科技有限公司
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