Method for preparing placenta hematopoietic stem cell preparation
A technology of hematopoietic stem cells and preparations, applied in the field of biomedicine, can solve the problems of low cell separation efficiency and inability to use directly, and achieve the effect of improving separation efficiency
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[0015] According to the present invention, wherein, according to a particularly preferred embodiment of the present invention, the enzyme solution contains 200-300 IU / mL type I collagenase, 30-50 IU / mL DNase I, 1.5-3.5 IU / mL dispase and 600-800IU / mL hyaluronidase; the freezing liquid contains 35-50mL / L dimethyl sulfoxide, 10-20g / L hydroxyethyl starch, 50-100g / L Human serum albumin, 10-20g / L dextran, 5-5.5g / L sodium chloride, 4.5-5g / L sodium gluconate, 3.4-3.9g / L sodium acetate, 0.3-0.45g / L Potassium chloride and 0.2-0.4g / L magnesium chloride. In this preferred embodiment, the present invention can further improve the separation efficiency of placental cells, and improve the hematopoietic colonization ability of frozen placental hematopoietic stem cells after returning to normal temperature.
[0016] According to the present invention, wherein, the weight ratio of the placental tissue and the enzyme solution can be 1: (0.5-5), preferably 1: (1-4); The volume ratio of the solu...
Embodiment 1
[0028] Digest the placental tissue with an enzyme solution at a temperature of 37°C for 30 minutes to obtain a digested product and filter through double-layer gauze to remove tissue residues in the digested product to obtain an enzymatic hydrolyzate; mix the enzymatically hydrolyzed solution with hydroxyethyl starch solution And let it stand for 20 minutes to separate the mixed materials to obtain the upper nucleated cell suspension; mix the nucleated cell suspension with the cryopreservation solution to obtain the placental hematopoietic stem cell preparation of this embodiment.
[0029] Wherein, the enzyme solution contains 250IU / mL type I collagenase, 40IU / mL DNase I, 2.4IU / mL dispase and 700IU / mL hyaluronidase; L of dimethyl sulfoxide, 15g / L of hydroxyethyl starch, 80g / L of human serum albumin, 15g / L of dextran, 5.2g / L of sodium chloride, 4.8g / L of sodium gluconate, 3.7g / L of sodium acetate, 0.40g / L of potassium chloride and 0.3g / L of magnesium chloride. The weight ratio...
Embodiment 2
[0031] Digest the placental tissue with an enzyme solution at a temperature of 37°C for 30 minutes to obtain a digested product and filter through double-layer gauze to remove tissue residues in the digested product to obtain an enzymatic hydrolyzate; mix the enzymatically hydrolyzed solution with hydroxyethyl starch solution And let it stand for 20 minutes to separate the mixed materials to obtain the upper nucleated cell suspension; mix the nucleated cell suspension with the cryopreservation solution to obtain the placental hematopoietic stem cell preparation of this embodiment.
[0032] Wherein, the enzyme solution contains 200IU / mL type I collagenase, 50IU / mL DNase I, 1.5IU / mL dispase and 800IU / mL hyaluronidase; L of dimethyl sulfoxide, 10g / L of hydroxyethyl starch, 100g / L of human serum albumin, 20g / L of dextran, 5g / L of sodium chloride, 5.0g / L of sodium gluconate, 3.4 g / L sodium acetate, 0.30g / L potassium chloride and 0.4g / L magnesium chloride. The weight ratio of the p...
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