Medium freeze-drying powder production process for lymphocyte culture

A production process and technology of lymphocytes, applied in the field of medium freeze-dried powder production process, can solve the problems such as no medium freeze-drying process or method has been found, and achieve the goal of improving activity and stability, stable effect and reducing production cost Effect

Active Publication Date: 2013-08-07
广州白云山拜迪生物医药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there is no public report that provides a specific medium freeze-drying process or method

Method used

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  • Medium freeze-drying powder production process for lymphocyte culture
  • Medium freeze-drying powder production process for lymphocyte culture
  • Medium freeze-drying powder production process for lymphocyte culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Freeze-dried culture medium of the present invention and traditional liquid culture medium activity, stability study result comparison

[0035] 1. Prepare respectively the 5-fold concentrated culture solution and the traditional liquid medium for freeze-drying of the present invention in the order of the formula in Table 1. Among them, RPMI1640 dry powder is from GIBCO Company, and newborn calf serum is from Hangzhou Sijiqing Company. After each material was added, stir with a magnetic stirrer at 300 rpm for two hours at room temperature to dissolve.

[0036]

[0037] 2. Filter the two solutions separately: the dissolved solution is first clarified and pre-filtered with a filter with a pore size of 1 um, and then sterile-filtered with a filter with a pore size of 0.22 um. The filter used is a 5-inch cartridge filter, and the corresponding 1 um and 0.22 um filter elements are all made of PES; the filtration flow rate is set to 1 L / hr. The filtration steps for non-co...

Embodiment 2

[0065] Comparison of the effects of lymphocyte culture medium freeze-dried powder prepared by different formulas or freeze-drying processes

[0066] See Table 4 for the differences and culture results between the freeze-dried lymphocyte medium process of the present invention and other freeze-dried medium processes. Wherein the concrete process parameter of process 1-3 is the same as embodiment 1, only the ratio of nitrogen and oxygen is different; The gas that process 4 feeds is common air, and other process parameters are with embodiment 1; Process 5 is due to the needs of dissolving mannitol, needs The filling volume is added, so the process parameters of sublimation and secondary drying are adjusted accordingly, and the gas introduced is ordinary air.

[0067] It can be seen from Table 4 that the osmotic pressure value of the lyophilized powder reconstituted solution prepared by adding mannitol in process 5 has exceeded the upper limit of normal lymphocyte culture (290mOSM...

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Abstract

The invention discloses a medium freeze-drying powder production process for lymphocyte culture, which comprises following steps: (1) preparing medium stock solution; (2) filtering medium stock solution, freezing, drying and obtaining the freeze-drying powder; (3) adding mixing gas containing nitrogen and oxygen into the freeze-drying powder, and obtaining the product. According to the process provided by the invention, a mixing gas containing nitrogen and oxygen at a certain ratio is added in the freeze-drying stage after the secondary drying ends, thereby activity and stability of the medium freeze-drying powder are unexpectedly improved. After a period of time,the freeze-drying powder compound solution is used for blood lymphocyte culture and chromosome karyotype analysis, and the two key active indexes such as lymphocyte conversion rate and mitotic phase index remain stable and are substantially higher than those of the traditional liquid medium. The lymphocyte medium freeze-drying powder prepared by the process provided by the invention has the advantages of long preservation time, stable effect, convenient storage, carrying and transport, thereby the commercialization of the lymphocyte medium is realized.

Description

technical field [0001] The invention relates to a production process of freeze-dried medium powder for lymphocyte culture. Background technique [0002] The traditional medium used for the preparation of chromosomes of peripheral blood lymphocytes is generally formed by adding a certain amount of serum and phytohemagglutinin (PHA) on the basis of liquid basal medium (such as RPMI1640). Use PHA to be in G 0 Stimulation of lymphocytes in the first stage, so that they enter the division cycle, so that the chromosome karyotype analysis can be carried out. However, lymphocyte culture medium in a liquid state is not conducive to storage at room temperature, and the active substances contained in it, such as glutamine and PHA, are easily inactivated. Even if stored at a low temperature of 2-8°C, the number of lymphocytes in the mitotic phase that can be analyzed for karyotype in the culture medium after a period of time decreases significantly. [0003] Utilizing freeze-drying t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0781C12N5/0783
Inventor 韦剑黄明陈建农王灿顶邱壮伟丘力功符美娟蔡小杰
Owner 广州白云山拜迪生物医药有限公司
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