CLINIC COMPLIANT METHOD FOR BANKING HUMAN PLACENTAL MESENCHYMAL CELLS
First Claim
1. A method for processing human placental cell sample, characterized in that said method comprises steps as follows:
- a. collecting human term placenta tissue, and protecting the tissue in DMEM containing 0.5% to 5%, preferably 1% of human cord blood serum;
b. isolating human placental amniotic and chorionic mersenchymal stromal cells from the placenta tissue obtained in step a;
c. in vitro expanding said human placental cells in a cell culture system that is free of any component of animal origin, and preferably said culture system is a DMEM-based medium, which further includes;
1). 5%-30%, preferably 10%-20% of human cord blood serum; and
2). 1% of penicillin/streptomycin solution;
d. determining antigen type (HLA-typing) of major histocompatibility (MHC) of the placental cells from each cell donor;
e. bar-coding the various HLA-typed cells obtained in step d and integrating HLA type information to registry information data for each cell donor;
f. preserving said placental mersenchymal stromal cells in cryopreservating solution and storing said cells in liquid nitrogen, and preferably, said cryopreservating solution is consisted of 50% human cord blood serum or autologous cord blood serum, 40% DMEM and 10% dimethyl sulphoxide (DMSO).
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Abstract
The present invention relates to a method for processing human placental cell sample, a human placental cell sample obtained according to said method for processing human placental cell sample, a human placental cell bank, a method for banking human placental cells, a method for searching human placental cell sample in said human placental cell bank according to the present invention, a method for preparing human cord blood serum, use of human placental cells obtained by said method for processing human placental cell sample or human placental cell bank established by said method for banking human placental cells in treating human dysfunction and diseases due to cell injury or cell malfunction, as well as a method for treating human dysfunction and diseases due to cell injury or cell malfunction.
29 Citations
11 Claims
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1. A method for processing human placental cell sample, characterized in that said method comprises steps as follows:
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a. collecting human term placenta tissue, and protecting the tissue in DMEM containing 0.5% to 5%, preferably 1% of human cord blood serum; b. isolating human placental amniotic and chorionic mersenchymal stromal cells from the placenta tissue obtained in step a; c. in vitro expanding said human placental cells in a cell culture system that is free of any component of animal origin, and preferably said culture system is a DMEM-based medium, which further includes; 1). 5%-30%, preferably 10%-20% of human cord blood serum; and 2). 1% of penicillin/streptomycin solution; d. determining antigen type (HLA-typing) of major histocompatibility (MHC) of the placental cells from each cell donor; e. bar-coding the various HLA-typed cells obtained in step d and integrating HLA type information to registry information data for each cell donor; f. preserving said placental mersenchymal stromal cells in cryopreservating solution and storing said cells in liquid nitrogen, and preferably, said cryopreservating solution is consisted of 50% human cord blood serum or autologous cord blood serum, 40% DMEM and 10% dimethyl sulphoxide (DMSO). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 10, 11)
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9. A method for preparing autologous cord blood serum, and preferably said autologous cord blood serum is used as a component of medium for expanding placental mersenchymal stromal cells, wherein said method comprises steps as follows:
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a. inserting the needle of a clinic syringe into the umbilical vein at the time of birth and taking the cord blood from the vein into the syringe; b. transferring the blood to a 50 ml centrifuge tube that is free of anticoagulants; c. allowing the blood to clot at 37°
C. for 30 to 60 minutes;d. cooling the clotted blood at 0 to 5°
C. for 15 to 45 minutes;e. having the blood centrifuged at 1000 g for 10 minutes; and f. transferring the serum to a collecting tube and incubating the serum at 50 to 56 for 30 minutes.
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Specification