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Stem cell spheroid induction culture solution and stem cell spheroid culture method

A culture medium and stem cell technology, applied in the field of cell culture medium

Inactive Publication Date: 2013-09-11
TIANJIN PEOPLE HOSPITAL
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  • Claims
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AI Technical Summary

Problems solved by technology

So far, there have been no reports in the literature on the use of culture medium to induce cells to produce stem cell pellets in an in vitro culture environment

Method used

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  • Stem cell spheroid induction culture solution and stem cell spheroid culture method
  • Stem cell spheroid induction culture solution and stem cell spheroid culture method
  • Stem cell spheroid induction culture solution and stem cell spheroid culture method

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Embodiment Construction

[0042] The present invention will be described in detail below in conjunction with the accompanying drawings and embodiments.

[0043] One. A kind of stem cell bead induction medium, it is made by the raw material of following proportioning, cobalt chloride (Cobalt chloride, CoCl 2 ) 142.758-285.516 mg, paclitaxel 2.135-4.27μg, fetal bovine serum 100-200ml, double antibiotics 10ml, mixed in EMEM medium at room temperature to 1000ml, filtered through a 0.2μ filter, and sterilized.

[0044] The stem cell bead induction culture medium is made according to the following ratio of raw materials: 214.137 mg of cobalt chloride, cobalt chloride, 2.989 μg of paclitaxel, 100 ml of fetal bovine serum, and 10 ml of double antibiotics are mixed in EMEM at room temperature for cultivation Base to 1000ml, filtered through a 0.2μ filter and sterilized.

[0045] 2. Culture method of stem cell pellets

[0046] 1. Culture of breast cancer cell line MDA-MB-231 cell stem cell pellets or BT-549 ce...

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Abstract

The invention discloses a stem cell spheroid induction culture solution and a stem cell spheroid culture method and belongs to cell culture solutions. The method comprises the steps of enabling cells cultured in an original culture solution to grow to reach 90% in fusion degree, using a mixed culture solution to culture the cells, performing primary anoxia culture in a carbon dioxide culture box until 90% of the cells die; enabling remaining adherent cells to be subjected to phosphate buffered saline (PBS) washing and be added into the original culture solution to continuously being cultured; after the cell density reaches 90%, replacing the original culture solution with the mixed culture solution again, performing secondary anoxia culture, and collecting suspended stem cell spheroids. Besides, the cells are induced in an in vitro environment to continuously generate plenty of stem cell spheroids, and the stem cell spheroids can express specific stem cell markers such as CD44, CD133, OCT3 / 4, Nanog, SOX-2 and ABCG2 which are normal or tumor stem cell markers.

Description

technical field [0001] The present invention relates to a cell culture solution, in particular to a stem cell spheroid that can induce cells to continuously produce a large number of stem cell spheroids (spheroids) by culturing cells (including tumor cell lines, immortalized cell lines and normal cell lines, etc.) sphere induction medium. Background technique [0002] Stem cells—especially cancer stem cells have become a hot spot in global tumor research. At present, the sorting of stem cells is basically carried out by using flow cytometry combined with stem cell-related surface markers, but stem cells are very easy to differentiate in an in vitro culture environment. In an in vitro culture environment, stem cells are more clearly characterized by the expression of stem cell markers and the ability to form stem cell pellets. Stem cell pellets are composed of a heterogeneous cell population: including primitive stem cells, progenitor cells and some differentiated cells. S...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095
Inventor 张诗武刘筠王弋嘉
Owner TIANJIN PEOPLE HOSPITAL
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