CN115261323A - Method for removing bacteria in umbilical cord blood cells - Google Patents
Method for removing bacteria in umbilical cord blood cells Download PDFInfo
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- CN115261323A CN115261323A CN202210930118.9A CN202210930118A CN115261323A CN 115261323 A CN115261323 A CN 115261323A CN 202210930118 A CN202210930118 A CN 202210930118A CN 115261323 A CN115261323 A CN 115261323A
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- 210000004027 cell Anatomy 0.000 title claims abstract description 57
- 210000004700 fetal blood Anatomy 0.000 title claims abstract description 40
- 241000894006 Bacteria Species 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000005406 washing Methods 0.000 claims abstract description 30
- 210000005087 mononuclear cell Anatomy 0.000 claims abstract description 21
- 210000003958 hematopoietic stem cell Anatomy 0.000 claims abstract description 18
- 230000003115 biocidal effect Effects 0.000 claims abstract description 14
- 229960000484 ceftazidime Drugs 0.000 claims abstract description 14
- NMVPEQXCMGEDNH-TZVUEUGBSA-N ceftazidime pentahydrate Chemical compound O.O.O.O.O.S([C@@H]1[C@@H](C(N1C=1C([O-])=O)=O)NC(=O)\C(=N/OC(C)(C)C(O)=O)C=2N=C(N)SC=2)CC=1C[N+]1=CC=CC=C1 NMVPEQXCMGEDNH-TZVUEUGBSA-N 0.000 claims abstract description 14
- 229960000282 metronidazole Drugs 0.000 claims abstract description 13
- VAOCPAMSLUNLGC-UHFFFAOYSA-N metronidazole Chemical compound CC1=NC=C([N+]([O-])=O)N1CCO VAOCPAMSLUNLGC-UHFFFAOYSA-N 0.000 claims abstract description 13
- 238000005119 centrifugation Methods 0.000 claims abstract description 9
- 238000005070 sampling Methods 0.000 claims abstract description 9
- 239000003242 anti bacterial agent Substances 0.000 claims abstract description 8
- 229940088710 antibiotic agent Drugs 0.000 claims abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 6
- 238000010257 thawing Methods 0.000 claims abstract description 3
- 210000004369 blood Anatomy 0.000 claims description 33
- 239000008280 blood Substances 0.000 claims description 33
- 238000012258 culturing Methods 0.000 claims description 13
- 230000022131 cell cycle Effects 0.000 claims description 11
- 239000006228 supernatant Substances 0.000 claims description 11
- 230000003203 everyday effect Effects 0.000 claims description 10
- 210000003743 erythrocyte Anatomy 0.000 claims description 9
- 238000003794 Gram staining Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 239000006916 nutrient agar Substances 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 6
- 239000008055 phosphate buffer solution Substances 0.000 claims description 6
- 239000002504 physiological saline solution Substances 0.000 claims description 6
- 238000001514 detection method Methods 0.000 claims description 5
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 230000002354 daily effect Effects 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 3
- 238000002372 labelling Methods 0.000 claims description 3
- 239000013642 negative control Substances 0.000 claims description 3
- 239000013641 positive control Substances 0.000 claims description 3
- 241001148471 unidentified anaerobic bacterium Species 0.000 abstract description 4
- 238000009640 blood culture Methods 0.000 abstract description 3
- 241001148470 aerobic bacillus Species 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000003501 co-culture Methods 0.000 abstract description 2
- 238000007865 diluting Methods 0.000 abstract description 2
- 230000001954 sterilising effect Effects 0.000 abstract description 2
- 238000011109 contamination Methods 0.000 description 6
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- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 210000003954 umbilical cord Anatomy 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
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- 210000002865 immune cell Anatomy 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 210000002826 placenta Anatomy 0.000 description 2
- 206010001986 Amoebic dysentery Diseases 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- 208000027205 Congenital disease Diseases 0.000 description 1
- 208000034423 Delivery Diseases 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 241000186394 Eubacterium Species 0.000 description 1
- 108090000279 Peptidyltransferases Proteins 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000011134 hematopoietic stem cell transplantation Methods 0.000 description 1
- 210000000777 hematopoietic system Anatomy 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000002906 medical waste Substances 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 210000005000 reproductive tract Anatomy 0.000 description 1
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- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
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Abstract
The invention discloses a method for removing bacteria in umbilical cord blood cells, which comprises the steps of cord blood hematopoietic stem cell thawing, washing centrifugation, mononuclear cell acquisition, secondary washing, cell and antibiotic coculture and identification. The beneficial effects of the invention are: diluting, washing, and adding conventional dosage of antibiotics in secondary washing for pretreatment; in the sterilization treatment, four groups of antibiotic concentrations were set for co-culture with the cells. The washing centrifugal blood culture sampling result shows that the concentration of bacteria is gradually reduced along with multiple times of washing; the identification results show that the best combination of four groups of antibiotic concentrations: 50mg/L of ceftazidime and 100mg/L of metronidazole infected by anaerobic bacteria, and the action time is 7 days; 100mg/L of ceftazidime and 12.5mg/L of metronidazole infected by aerobic bacteria are applied for 10 days.
Description
Technical Field
The invention relates to a method for rescuing umbilical cord blood cells, in particular to a method for removing bacteria in umbilical cord blood cells, belonging to the technical field of cell biology.
Background
The umbilical cord blood is blood remained in a placenta and an umbilical cord after delivery, ligation and separation of the umbilical cord, is rich in various cells, can be used for reconstructing a hematopoietic system and an immune system of a human body and treating diseases of the blood system, the immune system, genetic metabolism and congenital diseases, and is a very important biological genetic resource for human beings. As the umbilical cord and the placenta pass through the birth canal when the newborn is born, according to statistics, the microbial culture is carried out after the collection of the umbilical cord blood, the infection rate reaches 7-8 percent in a normal delivery mode, the caesarean section delivery rate is about 1 percent, and the average is about 4 percent. The contaminated umbilical cord blood mainly contains anaerobic bacteria, and the flora is mainly concentrated in enterobacteriaceae, staphylococcus, bacteroides and eubacterium, and is a normal flora of intestinal tract and reproductive tract. The treatment of the contaminated umbilical cord blood according to the medical waste is the final outcome in the past, and is a waste of biological resources from the viewpoint of genetic resources.
According to current statistical cord blood contamination data, about 4% of neonatal parents are unable to preserve neonatal cells for health care in newborns. If the antibiotic can be used properly, the cells of the microorganism-free newborn are again detected and stored by a cell washing method. From a certain point of view, this is a worthwhile endeavor. Ceftazidime can not synthesize cell walls by inhibiting transpeptidase from synthesizing transpeptidation in the bacterial cell walls, so that bacterial lysis is caused and the ceftazidime dies; has proven effective against gram-negative, gram-positive, anaerobic bacteria. Metronidazole is used to treat intestinal and extra-intestinal amebiasis and is currently widely used in the treatment of anaerobic infections.
Common types of biological contamination in cells are bacterial contamination, mycoplasma contamination, fungal contamination, and viral contamination. For umbilical cord blood, the extracted hematopoietic stem cells are used for treating immune cell diseases and blood system diseases, and the quality requirements of the hematopoietic stem cells are that the hematopoietic stem cells are free from bacteria, viruses, mycoplasma and fungi; therefore, the umbilical cord blood surface specimen infected with virus, mycoplasma and fungi is directly shielded according to the requirements of relevant specifications of blood station and hematopoietic stem cell transplantation, particularly virus infection; mycoplasma and fungi are not of interest for the present invention; however, cord blood contaminated with bacteria is an essential biological resource if bacteria can be removed by a certain means, and it is either used as a graft of hematopoietic stem cells or used as a natural killer cell, CAR-T cell, or the like by isolating leukocytes and culturing them in a targeted manner.
Disclosure of Invention
The present invention is directed to a method for removing bacteria from umbilical cord blood cells to solve at least one of the above problems.
The invention realizes the purpose through the following technical scheme: a method for removing bacteria from umbilical cord blood cells comprises the following steps
Step one, cord blood hematopoietic stem cell re-fusion: extracting and selecting the polluted umbilical cord blood hematopoietic stem cells, rapidly recovering the cells in a constant-temperature water bath kettle at 37 ℃, carrying out surface disinfection by using 75% alcohol, and placing the cells on a clean bench for subsequent treatment;
step two, washing and centrifuging: transferring the blood into a centrifuge tube, adding normal saline into the blood, and washing the umbilical cord blood hematopoietic stem cells; removing red blood cells (which are frozen and then recovered and then broken) through centrifugation, and extracting blood from the supernatant for culture;
step three, obtaining mononuclear cells: after centrifugation in the second step, the blood is divided into two layers, wherein the upper layer is red clear liquid, and the lower layer is mononuclear cells;
step four, secondary washing: adding a phosphate buffer solution containing ceftazidime and metronidazole at a certain concentration into the mononuclear cells obtained in the third step, and washing the mononuclear cells; centrifuging, separating into two layers, collecting upper layer transparent liquid and lower layer mononuclear cell, and collecting supernatant, and culturing;
step five, co-culturing the cells and antibiotics: grouping according to antibiotic addition concentration setting, and placing an addition culture medium in a CO2 incubator every day to observe the removal condition of bacteria and the growth condition of cells;
step six, identification: sampling every day, culturing by nutrient agar and performing gram staining for bacteria removal and identification; and detecting the cell cycle by using a flow cytometer for cell identification.
As a still further scheme of the invention: in the first step, the cord blood resuscitation requires the complete blood re-fusion.
As a still further scheme of the invention: in the second step, 0.9% physiological saline is used for diluting the blood, the amount of the physiological saline is 1/3 of the volume of the blood, and the blood is centrifuged for 1800prm/10min to remove hemolyzed red blood cells in the blood.
As a still further scheme of the invention: in the fourth step, phosphate buffer solution is used for washing cells for the second time, ceftazidime and metronidazole with antibiotic concentration of 100mg/L and 50mg/L are added, mixed uniformly and stood for 30min at room temperature; the mixture was centrifuged at 2000rpm/5min and the supernatant was discarded.
As a still further scheme of the invention: in the fifth step, the biological concentration is divided into four groups, and a negative control group and a positive control group are additionally arranged; GT-561 medium was added to each medium, and the growth state of the cells and the removal of bacteria were observed daily under an inverted microscope.
As a still further scheme of the invention: in the sixth step, sampling every day, performing gram staining, and detecting the bacteria removal condition by using a nutrient agar plate; and extracting each group of cells for cell cycle detection, labeling the cells with Propidium Iodide (PI) for analysis, and performing cell cycle analysis by a flow cytometer.
The invention has the beneficial effects that: the invention is diluted, washed, and the routine dosage of antibiotics is added for pretreatment in the secondary washing; in the sterilization treatment, four groups of antibiotic concentrations were set for co-culture with the cells. The washing centrifugal blood culture sampling result shows that the concentration of bacteria is gradually reduced along with multiple times of washing; the identification results show that the four groups of antibiotics have the best combination of concentrations: 50mg/L of ceftazidime and 100mg/L of metronidazole infected by anaerobic bacteria, and the action time is 7 days; 100mg/L of ceftazidime and 12.5mg/L of metronidazole infected by aerobic bacteria, and the action time is 10 days.
Drawings
FIG. 1 is a schematic view of the structure of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example one
As shown in FIG. 1, a method for removing bacteria from umbilical cord blood cells comprises the following steps
Step one, cord blood hematopoietic stem cell re-fusion: extracting and selecting polluted umbilical cord blood hematopoietic stem cells, rapidly recovering the cells in a constant-temperature water bath kettle at 37 ℃, carrying out surface disinfection by using 75% alcohol, and placing the cells on a superclean workbench for subsequent treatment;
step two, washing and centrifuging: transferring the blood into a centrifuge tube, adding normal saline into the blood, and washing the umbilical cord blood hematopoietic stem cells; removing red blood cells (the red blood cells are frozen and then recovered and then are crushed) through centrifugation, and extracting blood from the supernatant for culture;
step three, obtaining mononuclear cells: after centrifugation in the second step, the blood is divided into two layers, wherein the upper layer is red clear liquid, and the lower layer is mononuclear cells;
step four, secondary washing: adding a phosphate buffer solution containing ceftazidime and metronidazole at a certain concentration into the mononuclear cells obtained in the third step, and washing the mononuclear cells; centrifuging, separating into two layers, collecting upper layer transparent liquid and lower layer mononuclear cell, and collecting supernatant, and culturing;
step five, co-culturing the cells and antibiotics: grouping according to the antibiotic addition concentration, adding a culture medium into a CO2 incubator every day, and observing the removal condition of bacteria and the growth condition of cells;
step six, identification: sampling every day, culturing by nutrient agar and performing gram staining for bacteria removal and identification; and (3) detecting the cell cycle by using a flow cytometer for cell identification.
In the embodiment of the invention, in the first step, the cord blood resuscitation requires the complete blood thawing.
In the second step of the present invention, the blood is diluted with 0.9% physiological saline, the amount of the physiological saline is 1/3 of the volume of the blood, and the blood is centrifuged for 1800prm/10min to remove the hemolyzed erythrocytes.
In the fourth step of the invention, phosphate buffer is used for washing cells for the second time, ceftazidime and metronidazole are added with antibiotic concentration of 100mg/L and 50mg/L, and the mixture is uniformly mixed and stood for 30min at room temperature; the mixture was centrifuged at 2000rpm/5min and the supernatant was discarded.
Example two
As shown in FIG. 1, a method for removing bacteria from umbilical cord blood cells comprises the following steps
Step one, cord blood hematopoietic stem cell re-fusion: extracting and selecting polluted umbilical cord blood hematopoietic stem cells, rapidly recovering the cells in a constant-temperature water bath kettle at 37 ℃, carrying out surface disinfection by using 75% alcohol, and placing the cells on a superclean workbench for subsequent treatment;
step two, washing and centrifuging: transferring the blood into a centrifuge tube, adding normal saline into the blood, and washing the umbilical cord blood hematopoietic stem cells; removing red blood cells (the red blood cells are frozen and then recovered and then are crushed) through centrifugation, and extracting blood from the supernatant for culture;
step three, obtaining mononuclear cells: after centrifugation in the second step, the blood is divided into two layers, wherein the upper layer is red clear liquid, and the lower layer is mononuclear cells;
step four, secondary washing: adding a phosphate buffer solution containing ceftazidime and metronidazole at a certain concentration into the mononuclear cells obtained in the third step, and washing the mononuclear cells; centrifuging, separating into two layers, collecting upper layer transparent liquid and lower layer mononuclear cell, and collecting supernatant, and culturing;
step five, co-culturing the cells and antibiotics: grouping according to antibiotic addition concentration setting, and placing an addition culture medium in a CO2 incubator every day to observe the removal condition of bacteria and the growth condition of cells;
step six, identification: sampling every day, and performing nutrient agar culture and gram staining for bacteria removal and identification; and detecting the cell cycle by using a flow cytometer for cell identification.
In the embodiment of the invention, in the fifth step, four groups are carried out according to the antibiotic concentration, and a negative control group and a positive control group are additionally arranged; GT-561 medium was added to each medium, and the growth state of the cells and the removal of bacteria were observed daily under an inverted microscope.
In the embodiment of the invention, in the sixth step, sampling is carried out daily, gram staining and nutrient agar plate detection are carried out, and bacteria removal is carried out; and extracting each group of cells for cell cycle detection, analyzing the cells by Propidium Iodide (PI) labeling, and analyzing the cell cycle by a flow cytometer.
The working principle is as follows: through two antibacterial antibiotics of ceftazidime and metronidazole, by the methods of washing, centrifuging and co-culturing for many times, cord blood hematopoietic stem cells without bacteria are obtained again or cryopreserved or induced into immune cells by utilizing blood culture, gram staining and cell cycle detection.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (6)
1. A method for removing bacteria from umbilical cord blood cells, comprising: comprises the following steps
Step one, cord blood hematopoietic stem cell re-fusion: extracting and selecting polluted umbilical cord blood hematopoietic stem cells, rapidly recovering the cells in a constant-temperature water bath kettle at 37 ℃, carrying out surface disinfection by using 75% alcohol, and placing the cells on a superclean workbench for subsequent treatment;
step two, washing and centrifuging: transferring the blood into a centrifuge tube, adding normal saline into the blood, and washing the umbilical cord blood hematopoietic stem cells; removing red blood cells by centrifugation, and extracting blood from the supernatant for culture;
step three, obtaining mononuclear cells: after centrifugation in the second step, the blood is divided into two layers, wherein the upper layer is red clear liquid, and the lower layer is mononuclear cells;
step four, secondary washing: adding a phosphate buffer solution containing a certain concentration of ceftazidime and metronidazole into the mononuclear cells obtained in the third step, and washing the mononuclear cells; centrifuging, separating into two layers, collecting upper layer transparent liquid and lower layer mononuclear cell, and collecting supernatant, and culturing;
step five, co-culturing the cells and antibiotics: grouping according to antibiotic addition concentration setting, and placing an addition culture medium in a CO2 incubator every day to observe the removal condition of bacteria and the growth condition of cells;
step six, identification: sampling every day, culturing by nutrient agar and performing gram staining for bacteria removal and identification; and detecting the cell cycle by using a flow cytometer for cell identification.
2. The method for removing bacteria from umbilical cord blood cells as claimed in claim 1, wherein: in the first step, the cord blood resuscitation requires the complete blood thawing.
3. The method for removing bacteria from umbilical cord blood cells as claimed in claim 1, wherein: in the second step, 0.9% physiological saline is used to dilute the blood, the amount of the physiological saline is 1/3 of the volume of the blood, and the blood is centrifuged for 1800prm/10min to remove the hemolyzed red blood cells.
4. The method for removing bacteria from umbilical cord blood cells as claimed in claim 1, wherein: in the fourth step, phosphate buffer solution is used for washing cells for the second time, ceftazidime and metronidazole are added with antibiotic concentration of 100mg/L and mixed uniformly and stood for 30min at room temperature; the mixture was centrifuged at 2000rpm/5min and the supernatant was discarded.
5. The method for removing bacteria from umbilical cord blood cells as claimed in claim 1, wherein: in the fifth step, four groups are formed according to the antibiotic concentration, and a negative control group and a positive control group are additionally arranged; GT-561 medium was added to each medium, and the growth state of the cells and the removal of bacteria were observed daily under an inverted microscope.
6. The method for removing bacteria from umbilical cord blood cells as claimed in claim 1, wherein: sampling every day to perform gram staining and detecting the bacteria removal condition by a nutrient agar plate; and extracting each group of cells for cell cycle detection, labeling and analyzing the cells by using propidium iodide, and performing cell cycle analysis by using a flow cytometer.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0975430A (en) * | 1995-09-18 | 1997-03-25 | Masashi Funayama | Method for removing contaminant |
| CN105420191A (en) * | 2015-12-02 | 2016-03-23 | 上海华颜医药科技有限公司 | Preparing method for clinical cord blood monocyte rich in hematopoietic stem cells |
| CN107779425A (en) * | 2017-12-01 | 2018-03-09 | 重庆金时代生物技术有限公司 | A kind of suspended culture cell pollute after processing method |
| CN114058578A (en) * | 2021-12-29 | 2022-02-18 | 河北北科生物科技有限公司 | Method for amplifying NK cells from stored umbilical cord blood |
-
2022
- 2022-08-04 CN CN202210930118.9A patent/CN115261323A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0975430A (en) * | 1995-09-18 | 1997-03-25 | Masashi Funayama | Method for removing contaminant |
| CN105420191A (en) * | 2015-12-02 | 2016-03-23 | 上海华颜医药科技有限公司 | Preparing method for clinical cord blood monocyte rich in hematopoietic stem cells |
| CN107779425A (en) * | 2017-12-01 | 2018-03-09 | 重庆金时代生物技术有限公司 | A kind of suspended culture cell pollute after processing method |
| CN114058578A (en) * | 2021-12-29 | 2022-02-18 | 河北北科生物科技有限公司 | Method for amplifying NK cells from stored umbilical cord blood |
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Application publication date: 20221101 |