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Menstrual blood source stem cell preparation and preparation method and application thereof

A stem cell preparation and stem cell technology, applied in the field of biomedicine, to achieve the effects of uniform cell size, maintenance of activity and survival rate, and protection of activity

Active Publication Date: 2016-06-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on blood-derived stem cell preparations

Method used

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  • Menstrual blood source stem cell preparation and preparation method and application thereof
  • Menstrual blood source stem cell preparation and preparation method and application thereof
  • Menstrual blood source stem cell preparation and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Take the menstrual blood that has passed the inspection, dilute the menstrual blood with the same volume of PBS solution to form a menstrual blood dilution; take a separate centrifuge tube, add half the volume of the menstrual blood dilution Ficoll separation solution, slowly add the menstrual blood dilution, and then centrifuge at 700g for 15 minutes; the upper layer solution Move to another new centrifuge tube, wash with PBS twice, resuspend the pellet after centrifugation with serum-free medium, adjust the inoculation density to 7-8×10 5 / ml, inoculated into a petri dish, cross-mixed and placed in a carbon dioxide incubator for cultivation. Change the medium after 48 hours, change the medium after 72 hours, change the medium once every 3 days, and observe the formation of clones under the microscope around 7-14 days. When the cells grow to 70-80% confluent, trypsin digestion continues to cultivate.

[0042] Figure 1~2 The cell morphology of menstrual blood-derived ...

Embodiment 2～4

[0044] Table 1 Example 2-4 Menstrual Blood-derived Stem Cell Preparation Formula

[0045]

[0046] The menstrual blood-derived stem cells are resuspended after the albumin is mixed with physiological saline to obtain a menstrual blood-derived stem cell preparation.

Embodiment 5

[0051] Embodiment 5 preparation quality evaluation

[0052] Quality identification was carried out on the stem cell preparations prepared in Examples 2-4 and Comparative Examples 1-4. Each sample was checked 3 times. The method is:

[0053] After the preparation is made, the freshly prepared preparation is tested within 3 hours, and the detection content includes: stem cell morphology, survival rate, and the content of bacteria in the preparation. Among them, trypan blue staining was used for cell morphology detection and cell viability detection. The test results are shown in Table 3.

[0054] Test results within 3 hours after the preparation of table 3

[0055]

[0056] Note: A means that the cells have strong refraction, uniform size, and a spindle shape

[0057] B means that the cell volume increases, and there are abnormal shapes such as ellipse or irregular shape

[0058] --show not detected

[0059] The results show that: within 3 hours of preparation, in the ...

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Abstract

The invention relates to the field of biomedicine, in particular to a menstrual blood source stem cell preparation and a preparation method and application thereof. The menstrual blood source stem cell preparation comprises menstrual blood source stem cells, albumin and normal saline. The experiment indicates that the menstrual blood source stem cell preparation can keep the survival rate of the menstrual blood source stem cells at 99% or above, the cells are uniform in size, and the cells are each in a shuttle shape. After being preserved for 1 month at the temperature of 0-4 DEG C, the cells are still kept in good forms, the abnormal phenomenon such as increase of the size does not appear, the survival rate is still kept at 95% or above, and through surface antigen detection, the menstrual blood source stem cells still keep the characteristics of stem cells, no differentiation is generated, and good activity is achieved. The menstrual blood source stem cell preparation can achieve a good treatment effect when used for treating premature ovarian failure, and after the menstrual blood source stem cell preparation is used for a week, the number of follicles of a mouse is obviously increased.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a blood-derived stem cell preparation and its preparation method and application. Background technique [0002] Premature ovarian failure refers to a disease characterized by a decrease in serum estrogen levels and an increase in gonadotropin levels in women before the age of 40. According to epidemiological statistics, the incidence rate of women before the age of 40 is 1%, premature ovarian failure in primary menopause accounts for 10% to 28%, and premature ovarian failure in secondary amenorrhea accounts for 4% to 18%. There is an increasing trend year by year. Premature ovarian failure has become one of the main causes of infertility in women today, seriously affecting the quality of life of patients and increasing social and economic burdens. The etiology and pathogenesis of premature ovarian failure are complex and not yet fully understood. The treatment of premature ovarian f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K47/42A61P15/08A01N1/02
CPCA01N1/0226A61K35/28A61K47/42
Inventor 陈海佳王一飞葛啸虎马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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