CN105126083B - A kind of megakaryoblast preparation and its application, preparation method - Google Patents
A kind of megakaryoblast preparation and its application, preparation method Download PDFInfo
- Publication number
- CN105126083B CN105126083B CN201510541192.1A CN201510541192A CN105126083B CN 105126083 B CN105126083 B CN 105126083B CN 201510541192 A CN201510541192 A CN 201510541192A CN 105126083 B CN105126083 B CN 105126083B
- Authority
- CN
- China
- Prior art keywords
- preparation
- megakaryoblast
- stem cell
- cell
- platelet
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to stem cell medicine technical field, in particular to a kind of megakaryoblast preparation and its application, preparation method.The megakaryoblast preparation includes megakaryoblast, human serum albumin, stem cell factor, interleukin 2 and Sofflower injection.Megakaryoblast preparation of the present invention has significant improvement result for aleucia, after the patient's body platelet content after infusion restores normal level, can maintain longer period of time in vivo, using rear without any adverse reactions, using safe, and prepare it is simple, it is easy to use.
Description
Technical field
The present invention relates to stem cell medicine technical field, in particular to a kind of megakaryoblast preparation and its application, preparation
Method.
Background technique
Quantity is enough and normally functioning blood platelet is a ring indispensable in hemostasis, and the normal value of blood platelet is about
It is 150~450 × 103/mm3, therefore in a broad sense, it just can be described as aleukia (Thrombocytopenia) less than 150,000.
Foreign countries once directly took Cord blood from 5600~9100 fetuses to examine platelet count, found the ratio of newborn's aleukia
Example about 0.7~0.9% (Sola et al., 2000).Aleucia is also most normal in neonatal intensive care unit (NICU)
The hemorrhagic disease seen.Nowadays, hematopoietic stem cell transplantation is widely used to pernicious clinical treatment and non-malignant hematological disorder, entity
Tumor, hereditary metabolic disorders, autoimmune disease etc..In hematopoietic stem cell transplantation, the hemopoietic system of receptor is preprocessed
After be badly damaged, aleucia occurs for bone marrow suppression, decrease of platelet, and its recoverys is slower, easily leads to bleeding or even extremely
It dies.
However the treatment for aleucia, mainly direct feedback blood platelet, blood platelet is collected concentration and is got up,
Then cooperate physiological saline, be fed back to patient's body, to achieve the effect that quickly to supplement platelet counts.But feed back blood platelet
It is expensive, and platelet transfusion may face Inefficacy of Platelets Transfusion repeatedly, in addition, the stimulation of some thrombocytopoiesis because
Son such as thrombopoietin (TPO), may make patient the side effects such as myelofibrosis and myelosis sexual disorder occur.
Megacaryocyte (Megakaryocyte) is the cell that one of marrow is differentiated from candidate stem cell, is just
One of normal marrow can generate the mature cell of blood platelet, and predecessor is particle megacaryocyte.The cell volume is huge, mature
The rupture of megacaryocyte marginal portion forms blood platelet after falling off.Each megacaryocyte can averagely generate 2000 (200~7700
It is a) left and right blood platelet.Although megacaryocyte is that number is minimum in the hematopoietic cell of marrow, nucleated marrow cell population is only accounted for
0.05%, but its blood platelet generated is particularly important to the hemostatic function of body.Megacaryocyte is the Hematopoietic Stem from marrow
Cell differentiation development comes.Candidate stem cell breaks up first generates megakaryoblast, also referred to as macronucleus system colony forming unit
(colony forming unit-megakaryocyte, CFU-Meg).The endonuclear chromosome of progenitor stage is general
It is 2-3 times of body.When progenitor cells are 2 times of bodies or 4 times of bodies, cell has proliferative capacity, therefore this is that megakaryocytic series increases carefully
The stage of born of the same parents' quantity.When megakaryoblast is further divided into the megacaryocyte of 8-32 times of body, cytoplasm starts to break up, internal membrane system
It unites gradually complete.Finally there is a kind of film property substance that the cytoplasm of megacaryocyte is separated into many cells.When each cell is complete
Blood platelet is become when separating, blood platelet is fallen off by the gap between venous sinus sinusoid wall endothelium from megacaryocyte, into blood flow.
It has been reported that and is prepared into megakaryoblast injection, Neng Gougai with Cord blood megakaryoblast and human serum albumins
Decrease of platelet syndrome caused by kind chemicotherapy.But the therapeutic effect of the injection is undesirable, and easily sends out adverse reaction.Cause
This, a kind of therapeutic effect of urgent clinical needs is significant and the megakaryoblast preparation that has no adverse reaction.
Summary of the invention
In view of this, the present invention provides a kind of megakaryoblast preparation and its applications, preparation method.The megakaryoblast
Preparation has significant improvement result for aleucia, and the patient's body platelet content after infusion restores normal level
Afterwards, longer period of time can be maintained in vivo, and using rear without any adverse reactions, use is safe, and prepares simple, user
Just.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides a kind of megakaryoblast preparation, including megakaryoblast, human serum albumin, stem cell growth because
Son, interleukin 2 and Sofflower injection.
Megakaryoblast is that candidate stem cell is obtained by differentiation, also referred to as macronucleus system colony forming unit (colony
Forming unit-megakaryocyte, CFU-Meg).The endonuclear chromosome of progenitor stage is usually 2-3 times
Body.When progenitor cells are 2 times of bodies or 4 times of bodies, cell has proliferative capacity, therefore this is that megakaryocytic series increases cell quantity
Stage.When megakaryoblast is further divided into the megacaryocyte of 8-32 times of body, cytoplasm starts to break up, and endomembrane system is gradually complete
It is standby.Finally there is a kind of film property substance that the cytoplasm of megacaryocyte is separated into many cells.When each cell by it is completely separated when
As blood platelet, blood platelet is fallen off by the gap between venous sinus sinusoid wall endothelium from megacaryocyte, into blood flow.
Human serum albumin is people's blood product, and raw material comes from people's blood, caused by indication includes: 1. blood loss wounds, burn
Shock；2. tired pressure caused by brain edema and damage increases；3. oedema caused by cirrhosis and nephrosis or ascites；4. Hypoproteinemia
Prevention and treatment；5. hyperbilirubinemia of newborn；6. being controlled for cardiopulmonary bypass, the adjuvant treatment of burn, the auxiliary of haemodialysis
Treatment and adult respiratory distress syndrome (ARDS).
Stem cell factor (Stem Cell Growth Factors) is a kind of thin with stimulating self endogenous to do
Efficient protein matter, enzyme combination and the combination of the essential various Porcine HGFs of Stem Cell Culture In Vitro of intracellular growth,
Rich in epidermal growth factor (EGF), fibroblast growth factor (FGF), nerve growth factor (NGF), hepatocyte growth factor
(HGF), brain-derived neurotrophic factor (BDNF), platelet derived growth factor (PDGF), vascular endothelial growth factor
(VEGF), the multiple effective component such as transforming growth factor (TGF).Zoopery proves that stem cell factor can be activated quickly
The stem cell of suspend mode simultaneously promotes its growth, while microenvironment in adjustable body, provides advantageous growth conditions for stem cell.
Interleukin 2 is a kind of cell factor of chemotactic factor (CF) family, it is by many cells source (mainly by activating T
Cell generates), and the cell factor (mainly promoting lymphocyte growth, proliferation, differentiation) with polytropism effect；To body
Immune response and viral infection resisting etc. play an important role, and can stimulate by specific antigen or the T cell for causing mitogen factor to start
Proliferation；Energy activating T cell, promotes cell factor to generate；Stimulate NK cell Proliferation, enhance NK killing activity and generate cell because
Son, induction LAK cell generate；Promote B cell proliferation and secretory antibody；Activating macrophage.It can be used for clinical research and tumour
Treatment.
The main component of Sofflower injection is safflower, has effects that activating microcirculation and removing stasis medicinal, for treating occlusive cerebrovascular disease
Disease, coronary heart disease, vasculitis.It can be good at platelet aggregation-against, improve Platelet effect.
Megakaryoblast, human serum albumin, stem cell factor, leucocyte are situated between by megakaryoblast preparation of the present invention
After element -2 and Sofflower injection use in conjunction, by venous re-transfusion in vivo, it is circulated in hematological system, breaks up megakaryoblast,
Final differentiation thromboblast, and then important work is played by aleucia caused by Radiotherapy chemotherapy or other reasons
With, effect similar product significantly better than platelet transfusion and on the market, and it is without any side effects.
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
In some embodiments provided by the invention, include: in megakaryoblast preparation
In other embodiments provided by the invention, include: in megakaryoblast preparation
In other embodiments provided by the invention, include: in megakaryoblast preparation
Preferably, further including glucose injection in megakaryoblast preparation.The indication of glucose injection are as follows: mend
Fill energy and body fluid；It is lost (such as vomiting, diarrhea) for hypoalimentation caused by a variety of causes or a large amount of body fluid, it is complete intravenous
Nutrition, inanition ketosis；Hypoglycin；Potassemia；Hypertonic solution is used as tissue dewatering agent；Prepare peritoneal dialysis solution；Drug
Diluent；Vein method dextrose tolerance test；For preparing GIK (polarized solution).
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
In some embodiments provided by the invention, include: in megakaryoblast preparation
In other embodiments provided by the invention, include: in megakaryoblast preparation
In other embodiments provided by the invention, include: in megakaryoblast preparation
The present invention also provides application of the megakaryoblast preparation in preparation treatment aleukia disease drug；This is huge
Core ancestor cell preparation includes megakaryoblast, human serum albumin, stem cell factor, interleukin 2 and safflower injection
Liquid；Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
Preferably, further including glucose injection in megakaryoblast preparation；
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
The present invention also provides a kind of preparation methods of megakaryoblast preparation, comprising:
Megakaryoblast, human serum albumin, stem cell factor, interleukin 2 and Sofflower injection are mixed,
Obtain megakaryoblast preparation.
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
The present invention also provides a kind of preparation methods of megakaryoblast preparation, comprising:
By megakaryoblast, human serum albumin, stem cell factor, interleukin 2, Sofflower injection and grape
Sugared injection mixing, obtains megakaryoblast preparation.
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
The present invention also provides a kind of stem cell drugs, including megakaryoblast preparation provided by the invention；Macronucleus ancestral
Cell preparation includes megakaryoblast, human serum albumin, stem cell factor, interleukin 2 and Sofflower injection；Make
To be preferred, include: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
Preferably, further including glucose injection in megakaryoblast preparation；
Preferably, including: in megakaryoblast preparation
Preferably, include: in megakaryoblast preparation
The present invention provides a kind of megakaryoblast preparation and its applications, preparation method.The megakaryoblast preparation includes
Megakaryoblast, human serum albumin, stem cell factor, interleukin 2 and Sofflower injection.The present invention at least has
One of following advantage:
1, the patient's body platelet content after megakaryoblast preparation infusion of the present invention restores being averaged to normal level
Effective percentage is 92%, higher than the 85% of control group and 80%, shows megakaryoblast preparation of the invention for aleukia
Disease has significant improvement result；
It 2, can be in body after the patient's body platelet content after megakaryoblast preparation infusion of the present invention restores normal level
Interior maintenance longer period of time, hence it is evident that it is better than direct infusion blood platelet effect, also superior to existing megakaryoblast preparation effect；
3, megakaryoblast preparation of the present invention uses rear without any adverse reactions, and use is safe；
4, megakaryoblast preparation preparation of the present invention is simple, easy to use.
Specific embodiment
The invention discloses a kind of megakaryoblast preparation and its application, preparation method, those skilled in the art can be borrowed
Reflect present disclosure, is suitably modified realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are to this field
It is it will be apparent that they are considered as being included in the present invention for technical staff.Method and application of the invention has passed through
Preferred embodiment is described, related personnel obviously can not depart from the content of present invention, in spirit and scope to described herein
Methods and applications be modified or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
Raw materials used medicine or the city auxiliary material Jun Keyou in megakaryoblast preparation provided by the invention and its application, preparation method
Field is bought.The protein concentration 20% of human serum albumin, 10g/50mL；Stem cell factor concentration is 2 μ g/mL；Leucocyte is situated between
The concentration of element 2 is 20000IU/mL；Sofflower injection is 0.5g safflower crude drug in whole/mL.
Below with reference to embodiment, the present invention is further explained:
The preparation of 1 megakaryoblast of embodiment
1,0.5~1 × 10 are taken8The megakaryoblast of culture two weeks is transferred in 50mL centrifuge tube, and 300g is centrifuged 5min.
2, supernatant is abandoned, 10% glucose injection of 40mL is added into centrifuge tube and cleans cell, 200g is centrifuged 5min, clearly
It washes twice.
3, supernatant is abandoned, is resuspended 1 × 10 with 10% glucose injection of 80mL8Megakaryoblast cell, and add 2mL people
Blood albumin, 1.5mL stem cell factor, 0.5mL interleukin 2 and 11mL Sofflower injection.After mixing well, receive
It combines in 100mL infusion bag, it is spare.
Include: in the megakaryoblast preparation
The preparation of 2 megakaryoblast of embodiment
1,0.5~1 × 10 are taken8The megakaryoblast of culture two weeks is transferred in 50mL centrifuge tube, and 300g is centrifuged 5min.
2, supernatant is abandoned, 10% glucose injection of 40mL is added into centrifuge tube and cleans cell, 200g is centrifuged 5min, clearly
It washes twice.
3, supernatant is abandoned, is resuspended 1 × 10 with 10% glucose injection of 80mL8Megakaryoblast cell, and add 3mL people
Blood albumin, 1mL stem cell factor, 1mL interleukin 2 and 10mL Sofflower injection.After mixing well, it is collected in
It is spare in 100mL infusion bag.
Include: in the megakaryoblast preparation
The preparation of 3 megakaryoblast of embodiment
1,0.5~1 × 10 are taken8The megakaryoblast of culture two weeks is transferred in 50mL centrifuge tube, and 300g is centrifuged 5min.
2, supernatant is abandoned, 10% glucose injection of 40mL is added into centrifuge tube and cleans cell, 200g is centrifuged 5min, clearly
It washes twice.
3, supernatant is abandoned, is resuspended 1 × 10 with 10% glucose injection of 80mL8Megakaryoblast cell, and add 4mL people
Blood albumin, 0.5mL stem cell factor, 1.5mL interleukin 2 and 9mL Sofflower injection.After mixing well, receive
It combines in 100mL infusion bag, it is spare.
Include: in the megakaryoblast preparation
4 aleucia clinical trial of embodiment
1, test grouping:
Test group 1: it is treated using 1 preparation of embodiment；
Test group 2: it is treated using 2 preparation of embodiment；
Test group 3: it is treated using 3 preparation of embodiment；
Control group 1: being treated using megakaryoblast+human serum albumin, and said preparation is the preparation method comprises the following steps: use 80mL
10% glucose injection is resuspended 1 × 108Megakaryoblast cell, and 10mL human serum albumin is added, after mixing well, collect
It is spare in 100mL infusion bag；
Control group 2: infusion of therapeutic is carried out using platelet transfusion.
2, test method and result:
100 patients with aleucia are tested, are randomly divided into 5 groups, every group 20.By embodiment 1
~3 and the megakaryoblast preparation and platelet transfusion of control group be infused to 20 patient's veins of each group respectively, inspect periodically
Patient's body platelet content, and observe adverse reaction.Test result such as table 1.Calculate efficient, calculation method are as follows:
Effective percentage=platelet recovery is to normal level number of cases/test number of cases × 100%.
1 patient's body platelet content situation table of table
As can be seen from the above results, the patient's body platelet content after 1~3 preparation of embodiment of the present invention infusion
Restore to the average effectiveness level of normal level to be 92%, higher than the 85% of control group and 80%.
In addition, the patient's body platelet content after 1~3 preparation of embodiment of the present invention infusion restores normal level
Afterwards, longer period of time can be maintained in vivo, hence it is evident that it is better than direct infusion blood platelet effect, also superior to the macronucleus ancestral of control group 1
Cell preparation effect.And megakaryoblast preparation of the invention is using rear without any adverse reactions.Therefore macronucleus of the invention
Ancestor cell preparation has good improvement result for aleucia, and using safe.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (5)
1. a kind of megakaryoblast preparation, which is characterized in that by megakaryoblast, human serum albumin, stem cell factor, white
Cytokine -2, Sofflower injection and glucose injection composition；
Include: in the megakaryoblast preparation
2. megakaryoblast preparation according to claim 1, which is characterized in that include: in the megakaryoblast preparation
3. application of the megakaryoblast preparation as claimed in claim 1 or 2 in preparation treatment aleukia disease drug；Institute
It states in megakaryoblast preparation and includes:
4. a kind of preparation method of megakaryoblast preparation characterized by comprising
Megakaryoblast, human serum albumin, stem cell factor, interleukin 2, Sofflower injection and glucose are infused
Liquid mixing is penetrated, megakaryoblast preparation is obtained；Include: in the megakaryoblast preparation
5. a kind of stem cell drugs, which is characterized in that including megakaryoblast preparation as claimed in claim 1 or 2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510541192.1A CN105126083B (en) | 2015-08-27 | 2015-08-27 | A kind of megakaryoblast preparation and its application, preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510541192.1A CN105126083B (en) | 2015-08-27 | 2015-08-27 | A kind of megakaryoblast preparation and its application, preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105126083A CN105126083A (en) | 2015-12-09 |
| CN105126083B true CN105126083B (en) | 2019-02-19 |
Family
ID=54711892
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510541192.1A Active CN105126083B (en) | 2015-08-27 | 2015-08-27 | A kind of megakaryoblast preparation and its application, preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105126083B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101864396A (en) * | 2010-05-17 | 2010-10-20 | 中国人民解放军军事医学科学院野战输血研究所 | Method for inducing megakaryoblast and megakaryocyte in vitro |
-
2015
- 2015-08-27 CN CN201510541192.1A patent/CN105126083B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101864396A (en) * | 2010-05-17 | 2010-10-20 | 中国人民解放军军事医学科学院野战输血研究所 | Method for inducing megakaryoblast and megakaryocyte in vitro |
Non-Patent Citations (2)
| Title |
|---|
| 人胚胎干细胞的体外定向分化;刘卫东 等;《生命科学》;20071031;第19卷(第5期);第518-525 * |
| 红花注射液对血小板聚集、血液流变学及血栓形成的药效学研究;殷强;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20040615;E057-27 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105126083A (en) | 2015-12-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2009525297A (en) | Conditioned blood composition and method for producing the same | |
| Kim et al. | Dendritic cell dysfunction in patients with end-stage renal disease | |
| JP6995125B2 (en) | Methods and tools for making macrophages polarized to the M2 phenotype | |
| JP2019513154A (en) | Preparation of platelet pellet lysate and its use for the treatment of neurological disorders | |
| Hasiba-Pappas et al. | Platelet-rich plasma in plastic surgery: a systematic review | |
| CN115089755A (en) | Medical dressing composition of recombined fibronectin-humanized collagen with repairing effect and preparation method thereof | |
| US20240216441A1 (en) | Agent for improving cancer cachexia and method for improving cancer cachexia | |
| US20160089420A1 (en) | Activated leukocyte conditioned supernatant and uses for wound healing | |
| KR20120002530A (en) | Activated leukocyte composition | |
| CN105126083B (en) | A kind of megakaryoblast preparation and its application, preparation method | |
| CN111388649B (en) | Platelet-rich plasma placenta polypeptide composition and application thereof | |
| JP2001521528A (en) | Erythropoiesis stimulation method | |
| Matthews-Brzozowska et al. | Revitalization of facial skin based on preparations of patient own blood | |
| CN102212505A (en) | Immune killer cell, preparation method thereof, medicinal composition containing immune killer cell and set | |
| US11564943B2 (en) | Platelet concentrate for increase of cell regeneration and cell growth | |
| Karina et al. | Evaluation of plasma PDGF and VEGF levels after systemic administration of activated autologous platelet-rich plasma | |
| Yogev | A humoral solution: Autologous blood products and tissue repair | |
| WO2013036902A4 (en) | Submicron particles to decrease transfusion | |
| JP2023518085A (en) | Methods for making enhanced anti-inflammatory/anti-catabolic agents from autologous physiological fluids | |
| Bocci et al. | Ozone therapy in critical patients. Rationale of the therapy and proposed guidelines | |
| RU2619875C1 (en) | Method for oxidative stress correction under conditions of ultraviolet irradiation | |
| Elia | History of parenteral nutrition. | |
| Lever et al. | The proteins in Pemphigus Vulgaris: III. The effect of infusions of human serum albumin on the proteins in the blood serum of patients with Pemphigus Vulgaris | |
| CN113893330A (en) | Promoter cell gene serum factor for repairing organ skin | |
| JP3644690B2 (en) | Immunostimulator |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |