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Three-dimensional construction method for transfer liver cancer tissue models in vitro

A construction method and transferable technology, applied in the field of construction of three-dimensional metastatic liver cancer tissue in vitro model, can solve the problems of missing tumor cells, easy changes, inconsistent gene expression patterns, etc., to achieve easy repeatable research and simple application Effect

Inactive Publication Date: 2009-08-19
ZHONGSHAN HOSPITAL FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, limited by technology and methods, there is still a gap in the establishment of liver cancer tumor tissue models in vitro, and from the perspective of tumor cell biological characteristics, two-dimensional adherent cultured tumor cells in vitro for multiple passages are easy to change or lose some of the tumor cell expression. Type characteristics, biological characteristics, and the gene level expression pattern, drug screening situation is often inconsistent with clinical samples

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  • Three-dimensional construction method for transfer liver cancer tissue models in vitro
  • Three-dimensional construction method for transfer liver cancer tissue models in vitro
  • Three-dimensional construction method for transfer liver cancer tissue models in vitro

Examples

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Embodiment 1

[0028] A method for constructing a three-dimensional metastatic liver cancer tissue in vitro model, the specific steps are:

[0029] The first step: DMEM / F12 culture medium 1X (produced by GIBICO company), FBS (fetal bovine serum, Biointernational company) and penicillin and streptomycin solution (10000 units / ml penicillin, 10000 units / ml streptomycin are dissolved in 0.85g / 100ml sodium chloride solution, produced by GIBICO Company) is mixed in a volume ratio of 100:10:1 to obtain a mixed culture solution;

[0030] Step Two: Place the 1 x 10 7 Highly metastatic human liver cancer cells MHCC97H were suspended in 10ml of the mixed culture medium obtained in the first step, then placed in a 10ml RWV bioreactor (Synthecon, USA), and placed in a biodegradable scaffold polyglycolic acid-lactic acid copolymer Objects (length, width and height are 3mm, 3mm and 0.8mm respectively.), first stand still for 10min and then rotate at a speed of 7rpm / min for 10min, repeat three times, then...

Embodiment 2

[0032] A method for constructing a three-dimensional metastatic liver cancer tissue in vitro model, the specific steps are:

[0033] The first step: DMEM / F12 culture medium 1X (produced by GIBICO company), FBS (fetal bovine serum, Biointernational company) and penicillin and streptomycin solution (10000 units / ml penicillin, 10000 units / ml streptomycin are dissolved in 0.85g / 100ml sodium chloride solution, produced by GIBICO Company) is mixed in a volume ratio of 100:10:1 to obtain a mixed culture solution;

[0034] Step Two: Place the 1 x 10 7 Highly metastatic human liver cancer cells MHCC97H were suspended in 10ml of the mixed culture medium obtained in the first step, then placed in a 10ml RWV bioreactor (Synthecon, USA), and placed in a biodegradable scaffold polyglycolic acid-lactic acid copolymer Objects (length, width and height are 4mm, 4mm and 1.2mm respectively.), first stand still for 10min and then rotate at a speed of 8rpm / min for 15min, repeat three times, then...

Embodiment 3

[0036] A method for constructing a three-dimensional metastatic liver cancer tissue in vitro model, the specific steps are:

[0037] The first step: DMEM / F12 culture medium 1X (produced by GIBICO company), FBS (fetal bovine serum, Biointernational company) and penicillin and streptomycin solution (10000 units / ml penicillin, 10000 units / ml streptomycin are dissolved in 0.85g / 100ml sodium chloride solution, produced by GIBICO Company) is mixed in a volume ratio of 100:10:1 to obtain a mixed culture solution;

[0038] Step Two: Place the 1 x 10 7 Highly metastatic human liver cancer cells MHCC97H were suspended in 10ml of the mixed culture medium obtained in the first step, then placed in a 10ml RWV bioreactor (Synthecon, USA), and placed in a biodegradable scaffold polyglycolic acid-lactic acid copolymer (Length, width and height are respectively 4m, 4mm and 1mm.), first stand still for 10min and then rotate and cultivate with speed 7.6rpm / min for 10min, repeat three times, th...

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Abstract

The invention provides a method for constructing in-vitro model of 3D metastatic hepatoma-like tissues, characterized in that the method comprises the steps of continuously culturing highly metastatic human hepatoma cell line MHCC97H in a 3D rotary culture system combining an RWM bioreactor and a biologic scaffold PLGA (poly(lactic-co-glycolic acid)) for 15 days to form a metastatic hepatoma-like tissue model which has the characteristics closer to the in-vivo metastatic hepatoma tissues in terms of tissue morphological structure, specific gene expression and protein secretion, thereby better simulating and representing the growth characteristics and pathological changes of the in-vivo metastatic hepatoma solid tumor, therefore, the metastatic hepatoma-like tissue model is superior to the highly metastatic human hepatoma cell line model cultured by the tablet.

Description

technical field [0001] The invention relates to a method for constructing a three-dimensional metastatic liver cancer tissue in vitro model, and belongs to the technical field of in vitro tissue model construction methods. Background technique [0002] The traditional monolayer planar culture technology has been playing a leading role in the establishment of in vitro cancer cell models and the analysis of the biological behavior of cancer cells. However, limited by the cell culture method itself and the several passages of cancer cells, the morphology, polarity, and gene The expression and even the functional state are far from the actual pathophysiological conditions of cancer cells in vivo, and the resulting two-dimensional in vitro cancer cell models are increasingly challenged in the exploration of molecular pathological mechanisms of diseases. In recent years, the development of in vitro three-dimensional culture technology and the in-depth study of the biological chara...

Claims

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Application Information

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IPC IPC(8): C12N5/08C12N5/09
Inventor 崔杰峰刘银坤唐建华陈荣新徐长德康晓楠郭坤
Owner ZHONGSHAN HOSPITAL FUDAN UNIV
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