Method for preparing cord blood mononuclear cells

A nuclear cell and umbilical cord blood technology, applied in the cell-related field, can solve the problems of unguaranteed safety in premature infants, unusable application in premature infants, and low cell recovery rate, so as to facilitate infusion and use, avoid bacterial contamination, and cell recovery. active effect

Pending Publication Date: 2019-09-13
广州市天河诺亚生物工程有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The cell purity obtained by this method is better, the proportion of red blood cells is small, but the cell recovery rate is low, less than 50%
[0004] At the same time, the hydroxyethyl starch used in the centrifugal precipitation method of hydroxyethyl starch is a macromolecular substance, which is not easy to metabolize in the body, especially for premature infants.
Ficoll is also slightly toxic, and the safety of infusion in premature infants cannot be guaranteed

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  • Method for preparing cord blood mononuclear cells
  • Method for preparing cord blood mononuclear cells
  • Method for preparing cord blood mononuclear cells

Examples

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Embodiment 1

[0023] The preparation method of embodiment 1 umbilical cord blood mononuclear cells

[0024] (1) Add 28ml of anticoagulant to 27ml of cord blood collected, mix thoroughly and transfer to a sterile triple bag, in which the whole blood mononuclear cells (MNC) 10 6 / ml is 3.1, whole blood mononuclear cells (MNC) 10 8 / ml is 1.705;

[0025] (2) Centrifuge the transferred umbilical cord blood under the conditions of temperature 20°C, centrifugal force 500G, and centrifugation time 13 minutes;

[0026] (3) Put the centrifuged blood bag on the separation clamp to remove the plasma supernatant, and the precipitate is the mononuclear cell preparation. After centrifugal concentration, the umbilical cord blood mononuclear cell preparation is 10ml, and the whole blood mononuclear cell ( MNC)10 6 / ml is 16.28, whole blood mononuclear cells (MNC) 10 8 / ml is 1.628.

[0027] The only difference between Examples 2-30 and Example 1 is that the amount of cord blood collected is different,...

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Abstract

The invention provides a method for preparing cord blood mononuclear cells. The method comprises the following steps: (1) transferring collected cord blood into a sterile triple bag; (2) carrying outcentrifugal separation on the transferred cord blood; and (3) placing the centrifuged blood bag on a plasma separation clamp to remove plasma supernatant, and acquiring precipitate, namely a mononuclear cell preparation. The preparation method disclosed by the invention does not use hydroxyethyl starch and ficoll lymphocyte separation liquid, the prepared mononuclear cell preparation is safe and has no side effect, the recovery rate of mononuclear cells reaches more than 95%, and the cell activity is more than 93%. The invention also provides a cord blood mononuclear cell preparation. The preparation has small volume of just 10ml, and is convenient to infusion and use for newborn.

Description

technical field [0001] The invention relates to the field of cell technology, in particular to a method for preparing umbilical cord blood mononuclear cells. Background technique [0002] In immune cell therapy, peripheral blood mononuclear cells (PBMC) are generally separated by hydroxyethyl starch (HES) centrifugation or Ficoll density gradient centrifugation. [0003] Although the hydroxyethyl starch centrifugal precipitation method is relatively simple in operation, and the cell recovery rate can reach more than 80%, the effect of cell separation is not good, and the purity of the obtained cells is difficult to meet the requirements, and there are more platelets, red blood cells and granulocytes. Wait. Ficoll density gradient centrifugation method, due to the large proportion of red blood cells and granulocytes, they will sink to the bottom of the tube after centrifugation; while the specific gravity of lymphocytes and monocytes is less than or equal to the specific gra...

Claims

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Application Information

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IPC IPC(8): C12N5/078
CPCC12N5/0634C12N2509/00
Inventor 嵐山芮魏伟马天宝林汉标许超
Owner 广州市天河诺亚生物工程有限公司
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