Source of people iPS stem cell in vitro directed differentiation is the kit and method of cardiac muscle cell

Source of people iPS stem cell in vitro directed differentiation is the kit and method of cardiac muscle cell

  • CN 106,244,527 B
  • Filed: 08/29/2016
  • Issued: 10/08/2019
  • Est. Priority Date: 08/29/2016
  • Status: Active Grant
First Claim
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1. a kind of for that will induce multi-potent stem cell the kit that vitro directed differentiation is cardiac muscle cell, the kit includes theOne culture solution, first culture solution include basal medium DMEM/F12, further, first culture solution further include:

  • 4~11 μ

    g/ml of insulin;

    40~70mg/ml of 2- Ascorbic acid 2-phosphate;

    40~80 μ

    g/ml of sodium selenite;

    150~200 μ

    g/ml of mankind'"'"'s basic fibroblast growth factor;

    1 90~140 μ

    g/ml of human transforming growth factor TGF β



    30~60mg/ml of transferrins;

    NaHCO34~8wt%;

    Wherein first culture solution further include;

    1~3 μ

    M of TZV or 8~12 μ

    M of Y-27632;

    Wherein the pH value of first culture solution is less than 7;

    The kit further includes the second culture solution, and second culture solution includes CDM culture solution, further, described secondCulture solution further includes 8~12 μ

    M of CHIR99021;

    Wherein the CDM culture solution includes basal medium RPMI-1640,0.08~0.12vol%BSA, and 1 ×

    MTG, 200~250 μ

    g/mL magnesium ascorbyl phosphate;

    The kit further includes third culture solution, and the third culture solution includes CDM culture solution, further, the thirdCulture solution further includes the basic fibroblast growth factor of 4~6ng/ml;

    Wherein the CDM culture solution includes basal medium RPMI-1640,0.08~0.12vol%BSA, and 1 ×

    MTG, 200~250 μ

    g/mL magnesium ascorbyl phosphate;

    The kit further includes the 4th culture solution, and the 4th culture solution includes CDM culture solution and used thirdCulture solution, further, the 4th culture solution further include 2~8 μ

    M of IWP2/4, IWR-1 or 1~3 of 1~3 μ

    M μ

    MWnt-C59;

    Wherein, the CDM culture solution includes basal medium RPMI-1640,0.08~0.12vol%BSA, and 1 ×

    MTG, 200~250 μ

    g/mL magnesium ascorbyl phosphate;

    Wherein, the CDM culture solution and the volume ratio of used third culture solution be 0.5~2;

    The kit further includes the 5th culture solution, and the 5th culture solution is CDM culture solution or the 5th culture solution includesBasal medium RPMI-1640 further includes the B27 of the KOSR or 2~4vol% of 2~4vol%;

    Wherein the CDM culture solution includes basal medium RPMI-1640,0.08~0.12vol%BSA, and 1 ×

    MTG, 200~250 μ

    g/mL magnesium ascorbyl phosphate.

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