CN104862277B - High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid - Google Patents
High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid Download PDFInfo
- Publication number
- CN104862277B CN104862277B CN201410066574.9A CN201410066574A CN104862277B CN 104862277 B CN104862277 B CN 104862277B CN 201410066574 A CN201410066574 A CN 201410066574A CN 104862277 B CN104862277 B CN 104862277B
- Authority
- CN
- China
- Prior art keywords
- stem cell
- cell
- culture medium
- culture
- cell factor
- 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
Abstract
The present invention relates to high-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid.Disclose a kind of culture medium and cultural method of in vitro amplifying candidate stem cell, candidate stem cell can be made to carry out, effectively with a large amount of amplification, greatly improving the amplification in vitro efficiency of candidate stem cell on the basis of original dryness feature is kept by the culture medium and cultural method.
Description
Technical field
The invention belongs to biomedical sectors；More particularly it relates to high-efficiency in-vitro human hematopoietic stem cell amplification training
Nutrient solution formula.
Background technology
Stem cell (Stem Cells) is a kind of cell colony with self-renewing and multi-lineage potential, i.e., these are thin
Born of the same parents can maintain the size of own cells group by cell division, while can be further differentiated into again as a variety of different tissues
Cell, so as to which medical field is referred to as " omnipotent cell ".This unique characteristic becomes regenerative medicine and transplantation medicine
An indispensable cell derived.Also numerous diseases are made it, such as hematologic disease, Parkinson's disease, diabetes, spinal cord damage
The best candidate of the cell therapies such as wound, cancer and heart disease, thus for stem cell research in clinical practice field meaning
It is great.
Candidate stem cell (Hematopoietic Stem Cell, HSC), which is that a group being present in hematopoietic tissue is original, to be made
Haemocyte, it is not that tissue fixes cell, be may be present in hematopoietic tissue and blood.Candidate stem cell is in 2 Zhou Shike of Human embryo
Yolk bag is come across, for gestation after 5 months, marrow starts hematopoiesis, and marrow becomes the main source of candidate stem cell after birth.It is making
In haemal tissue, stem cell proportion is very few.In modern medicine, candidate stem cell has weight in terms of bone-marrow transplantation and disease treatment
It acts on.
Candidate stem cell in application process a bottleneck be can not effectively amplification in vitro and in vitro expand in
Its dryness can be lost.The cell of quantity available is considerably less, cannot meet clinical the needs of curing at all.Known candidate stem cell
Amplification method have method using feeder cells, the method cannot make sub- stem cell maintain the maturation identical with mother stem cell
State, most preferably can be 5 times or so with expanding stem cells, and introduce exogenous cell interference, are unfavorable for after amplification for clinic
Transplanting easily improves graft-versus-host reaction (GVHD) occurrence risk.
Therefore, there is an urgent need in the art to provide the reagent and method of the effective culture and amplification for being directed to candidate stem cell.
Invention content
The purpose of the present invention is to provide high-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid.
In the first aspect of the present invention, a kind of cell factor composition for hematopoietic stem cell expansion culture, institute are provided
The composition stated is cell factor composition 1, including：
In a preference, the cell factor composition 1 includes：
In another preferred example, the cell factor composition 1 includes：
In another aspect of this invention, the purposes of any cell factor composition 1 in front is provided, is used to prepare and makes
The culture medium of hemocytoblast amplification cultivation.
In another aspect of this invention, a kind of culture medium for hematopoietic stem cell expansion culture, the culture are provided
Base is culture medium 1, including：IMDM culture mediums；With any cell factor composition 1 in front.
In a preference, which further includes fetal calf serum.
In another preferred example, the fetal calf serum is the dosage according to volume 8-12%, preferably 10%.
In another aspect of this invention, the purposes of the culture medium 1 is provided, is made for amplification cultivation (for cultured in vitro)
Hemocytoblast.
In another preferred example, the candidate stem cell is human cord blood candidate stem cell.
In another aspect of this invention, a kind of kit for hematopoietic stem cell expansion culture, the reagent are provided
Box includes：Any cell factor composition 1 in front.
In another preferred example, it is further included in the kit：Cell factor composition 2, the cell factor composition 2 wrap
It includes：
In another preferred example, the cell factor composition 2 includes：
In another preferred example, it is further included in the kit：IMDM culture mediums；The cell factor composition 1
And/or cell factor composition 2 is divided in different containers from IMDM culture mediums and places or be formulated in IMDM culture mediums
In.
In another aspect of this invention, a kind of method of amplification cultivation candidate stem cell is provided, the method includes：
(1) application foregoing culture medium 1 culture CD34+ candidate stem cell, replaced during culture 1-2 times it is fresh
Culture medium 1 (preferably the 3rd ± 0.5 day after starting, replaces 1 fresh culture medium 1)；
(2) after culture starting the 6th ± 1 day (preferably 6 ± 0.5 days), culture medium is changed to culture medium 2, during culture
It replaces 1-2 fresh culture medium 2 and (preferably the 9th ± 1 day after starting (preferably 9 ± 0.5 days), replaces 1 fresh training
Support base 2)；
Wherein, the culture medium 2 includes IMDM culture mediums and cell factor composition 2, which wraps
It includes：
In another preferred example, the 12nd ± 1 day after culture originates, culture is terminated, harvests candidate stem cell.
The other aspects of the present invention are apparent to those skilled in the art due to this disclosure
's.
Description of the drawings
The total karyocyte expanding effect figure of different incubation times after Fig. 1, stem cell separating and purifying.
Different incubation time CD34+ cell expanding effect figures after Fig. 2, stem cell separating and purifying.
Fig. 3, combination of cytokines amplifying candidate stem cell microscope figure is used.
Specific embodiment
The technical issues of for candidate stem cell efficient amplification is difficult to realize in the prior art, the present inventor is by deep
Research, develops a kind of culture medium and cultural method of in vitro amplifying candidate stem cell, passes through the culture medium and cultural method
Candidate stem cell can be made to carry out, effectively with a large amount of amplification, it is thin greatly improving Hematopoietic Stem on the basis of original dryness feature is kept
The amplification in vitro efficiency of born of the same parents.
As used herein, it is " basic to include "comprising", " mainly by ... form and (be made) " for term " containing " or " comprising "
On by ... form " and " by ... form ".
Cell factor composition and culture medium
The present inventor is optimized for the cell factor of in vitro amplifying candidate stem cell.According to hematopoietic stem cell expansion not
The same stage provides different cell factor compositions, including cell factor composition 1 and cell factor composition 2.
The cell factor composition 1 includes：Stem cell factor (SCF), flt-L (FLT-3L), blood platelet because
Sub (TPO), neutrophil leucocyte colony stimulating factor (G-CSF), interleukin-13 (IL3), interleukin 6 (IL6) and Sall sample albumen 4B
(Sall4B).Above-mentioned each cell factor is added to suitable ratio in cell growth medium, can be provided for candidate stem cell
The culture medium of suitable isolated growth environment promotes the growth and amplification of candidate stem cell.As the preferred embodiment of the present invention, use
It is as shown in table 1 in the dosage of each component for the culture medium for preparing the present invention.
Table 1
| Content | Preferred amounts | More preferably amount | Most preferred amount | |
| Sall sample albumen 4B | 1-10ng/ml | 1.5-8ng/ml | 2-6ng/ml | 3ng/ml |
| Stem cell factor | 50-500ng/ml | 60-300ng/ml | 80-150ng/ml | 100ng/ml |
| Flt-L | 50-500ng/ml | 60-300ng/ml | 80-150ng/ml | 100ng/ml |
| Platelet factor | 5-100ng/ml | 6-70ng/ml | 8-30ng/ml | 20ng/ml |
| Neutrophil leucocyte colony stimulating factor | 5-100ng/ml | 6-70ng/ml | 8-30ng/ml | 12.5ng/ml |
| Interleukin-13 | 10-250ng/ml | 15-150ng/ml | 20-50ng/ml | 25ng/ml |
| Interleukin 6 | 5-100ng/ml | 6-70ng/ml | 8-30ng/ml | 12.5ng/ml |
The cell factor that table 1 is formulated is dissolved in cell culture medium, obtains culture medium 1, so as to be provided for candidate stem cell
Suitable growth and amplification environment.The cell culture medium can select IMDM culture mediums or similar cell culture medium.
The cell factor composition 2 includes：Sall sample albumen 4B, stem cell factor, flt-L, blood platelet because
Son.Above-mentioned each cell factor is added to suitable ratio in cell growth culture solution, can be candidate stem cell culture medium prescription
2.It is as shown in table 2 for preparing the dosage of each component of the culture medium of the present invention as the preferred embodiment of the present invention.
Table 2
| Content | Preferred amounts | More preferably amount | Most preferred amount | |
| Sall sample albumen 4B | 1-10ng/ml | 1.5-8ng/ml | 2-6ng/ml | 3ng/ml |
| Stem cell factor | 50-500ng/ml | 60-300ng/ml | 80-150ng/ml | 100ng/ml |
| Flt-L | 50-500ng/ml | 60-300ng/ml | 80-150ng/ml | 100ng/ml |
| Platelet factor | 3-50ng/ml | 5-30ng/ml | 6-20ng/ml | 10ng/ml |
The cell factor that table 2 is formulated is dissolved in cell culture medium, obtains culture medium 2, so as to be provided for candidate stem cell
The culture medium prescription of preferred simplification.
Above-mentioned culture medium prescription 1 and 2 after the present inventor optimizes contains enough and rational promotion cell growth and expansion
The composition of increasing is conducive to the culture of candidate stem cell.
It is people in the art for preparing Sall4B, SCF, TPO, Flt-3L, G-CSF, IL3 and IL6 of culture medium
What member was easily obtained, such as can be bought or can be obtained by artificial synthesized or recombinant expression by commercial sources.
Cultural method
The present invention also provides a kind of method of amplification cultivation candidate stem cell, the method includes：Using described thin
Intracellular cytokine composition is added in cell growth medium, cultivates candidate stem cell.Preferably, the method includes：(1) it applies
Culture medium 1 cultivates the candidate stem cell of CD34+, and 1-2 fresh culture medium 1 is replaced during culture；Preferably the 3rd after starting
± 0.5 day, replace 1 fresh culture medium 1；(2) the 6th ± 1 day after culture starting, culture medium is changed to culture medium 2, is cultivated
Period replace 1-2 fresh culture medium 2, preferably the 9th ± 1 day after starting, preferably 9 ± 0.5 days, replacement 1 time it is fresh
Culture medium 2.
The present invention is by candidate stem cell by the stem cell growth culture medium (culture medium 1 and culture medium 2) of optimization in external
Safe and efficient amplification not only can largely obtain the scientific research needs for stem-cell research laboratory, it is often more important that Ke Yiman
Sufficient stem cell transplantation in clinic and the needs of war preparedness storage.
In the clinical trial of this field, have existed some schemes and carry out amplifying candidate stem cell, but in the division of stem cell
Cheng Dangzhong, a daughter cell retains maturity state identical with female stem cell, but another the situation of differentiation then occurs, has deposited
Scheme can exactly make expansion of stem cells only a generation or two generations, under keeping the cell quantity of original dryness after this significantly
Drop.The present invention is matched using the amplifying candidate stem cell in vitro containing Sall4B, SCF, TPO, Flt-3L, G-CSF, IL3 and IL6
Side carrys out the technology of efficient amplification candidate stem cell, compared with traditional hematopoietic stem cell expansion method, has following several main
Feature and advantage：First, expression of exogenous genes is not introduced, therefore do not change the Genome stability of former stem cell, no tumorigenesis
Risk；Second, it is not related to stem cell transplantation safety problem caused by the exogenous cells such as feeder cells mix；Third, this
Invention can make to increase CD34+ stem/progenitor cells quantity close to 70 times in Stem cells cultured in vitro 9 days, and total karyocyte sum is being trained
Increase at least 160 times when supporting 9 days, total karyocyte can increase by hundreds times or more when cultivating 16 days.
Use its amplification efficiency of the culture medium expanding stem cells of in vitro stem cell invented reports to be 5 at present at present
Times or so, the expansion of stem cells for needing introducing expression of exogenous genes and carrying out can most preferably reach 160 times to 200 times of amplification
Effect, and dryness for candidate stem cell and gene stability cause interference.And the present invention is because having used special formulation
Culture amplifying candidate stem cell greatly improve amplification efficiency, with tradition culture formula compare have significantly higher expansion
Increasing Efficiency, and can be good at keeping the essential characteristics such as stem cells self-renewal and multipotency differentiation.
With reference to specific embodiment, the present invention is further explained.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.Test method without specific conditions in the following example, usually according to conventional strip
Part such as J. Pehanorm Brookers etc. are write, Molecular Cloning:A Laboratory guide, the third edition, Science Press, condition described in 2002 or
According to the normal condition proposed by manufacturer.
Embodiment 1, formula A joint formula A1 culture human hematopoietic stem cells
According to the amount for preparing 200ml amplification culture mediums, in the following order and following ingredients are shifted under super-clean bench：
Iscove’s Modified Dulbecco’s Medium(IMDM)(respectively containing 10% (v/v) fetal calf serum and
Without fetal calf serum) 200ml.100ml is taken out from the culture medium and adds in cell factor, makes cell factor final concentration of
IL325ng/ml, IL612.5ng/ml, TPO20ng/ml and G-CSF are 12.5ng/ml, and SCF 100ng/ml, FLT-3L are
100ng/ml, Sall4B3ng/ml (are formulated A as growth medium containing 10% fetal calf serum --- ---；Without tire ox blood
--- --- formula B clearly)；The conduct basis culture medium of remaining not factor-containing (is formulated C containing 10% fetal calf serum --- ---；No
Containing fetal calf serum, --- --- is formulated D).
Fresh Cord blood is taken, CD34+ cells is detached using Mei Tian Ni company MACS magnetic bead sortings system, obtains cell quantity
It is 0.36 × 106A cell, streaming identify that CD34+ cell colonys purity is 93.3%.The volume for calculating culture medium cultivating
Cell concentration in base is 5 × 103A cell/ml takes the cell of wherein half to add in culture medium prescription A36ml, and average mark
It is attached in six well culture plate of Corning Incorporated's low adsorption, 6 holes and cultivates, per hole 6ml culture medium As (or B) containing cell 3 × 104It is a, and
This culture plate is labeled as I plates；The cell of remaining half is separately added into culture medium prescription C and formula D and is similarly grasped
Make, labeled as П plates.Be placed in incubator with micro- sem observation and then by pipe (37 DEG C, 5%CO2)。
3rd day, culture plate is taken out from incubator, A is replaced per hole to I plates or B, 1200rpm are centrifuged 5 minutes, is collected
Cell precipitation is simultaneously resuspended in 6ml fresh cultures.C is replaced per hole to П plates or D, 1200rpm are centrifuged 5 minutes, is collected thin
Born of the same parents are precipitated and are resuspended in 6ml fresh cultures.Culture plate is placed in incubator again and is cultivated.
Again according to the amount for preparing 200ml amplification culture mediums, in the following order and following ingredients are shifted under super-clean bench：Training
Base Iscove ' s Modified Dulbecco ' s Medium200ml are supported, 100ml is taken out from the culture medium and is added in carefully
Intracellular cytokine makes cell factor final concentration of SCF100ng/ml, FLT-3L100ng/ml, TPO10ng/ml, Sall4B3ng/ml works
(A1 is formulated for growth medium containing fetal calf serum --- ---；Without fetal calf serum, --- --- is formulated B1)；It is remaining to be free of cell
The conduct basis culture medium of the factor (is formulated C containing fetal calf serum --- ---；Without fetal calf serum, --- --- is formulated D).
6th day, A1 or B1 culture mediums are replaced in the above described manner per hole to I plates.C or D trainings are replaced per hole to П plates
Support base.Culture plate is placed in incubator again.
It carries out within 9th day and same operation in the 6th day.
Amplification in 12nd day is completed.It is prepared in vitro in super-clean bench, under aseptic condition molten containing 2% (w/v) human albumin physiology
Liquid, as washing buffer.All holes containing amplifying cells are collected from incubator every hole is carried out visually to see with microscope
It examines.After observation, it will be transferred in 15ml centrifuge tubes after the suction uniformly of the content in every hole respectively, 1200rpm centrifugations 5 at room temperature
Minute.It adds in 5ml washing buffers and cell precipitation is resuspended, 1200rpm is centrifuged 5 minutes at room temperature again.Gained cell is resuspended
It precipitates and adds to 5ml washing buffers to form final pipe, according to the 1-6 holes of every piece of culture plate into line label.From it is described most
The counting that cell suspension carries out karyocyte total amount with flow cytometer is taken out in whole pipe.
Comprehensive analysis, such as Fig. 1-2, No. I culture plate prescription A are carried out to total cell group using flow cytometer：Always there is core
Cell is averaged amplification times as 354.1154, and the wherein CD34+ cells amplification times that be averaged are 75.65921 times；I culture plates are matched
Square B：Total karyocyte amplification times that be averaged are 85.84615, and the wherein CD34+ cells amplification times that are averaged are 12.09833 times.П
Number culture plate prescription C：Total karyocyte amplification times that be averaged are 12.11538, and the wherein CD34+ cells amplification times that are averaged are
2.988812 again；No. П culture plate prescription D：Total karyocyte amplification times that be averaged are 6.576923, and wherein CD34+ cells are averaged
Amplification times are 5.153649 times.Fig. 3 is to be formulated A, B, C, the candidate stem cell form that D was observed in the 6th day microscope respectively.
Thus, it could be seen that the application of formula A joint formulas A1, can effectively expand human hematopoietic stem cell.It is not added with fetal calf serum
It is formulated B and formula B1 use in conjunction, expanding effect is also ideal.
Embodiment 2, other formula culture human hematopoietic stem cells
Based on the cell factor, inventor has also prepared some other formula, with IMDM10% (v/v) fetal calf serum)
As cell culture medium, wherein being separately added into the cell factor being formulated as shown in table 3- tables 4.Obtain formula E/E1；It is formulated F/F1；
It is formulated G/G1；It is formulated H/H1.
Table 3,1 (unit of culture medium：ng/ml)
| It is formulated E | It is formulated F | It is formulated G | It is formulated H | |
| Sall4B | 6 | 3 | 3 | 3 |
| SCF | 200 | 150 | 150 | 100 |
| FLT-3L | 200 | 200 | 150 | 150 |
| TPO | 100 | 50 | 50 | 25 |
| G-CSF | 100 | 50 | 25 | 12.5 |
| IL3 | 150 | 100 | 50 | 25 |
| IL6 | 100 | 50 | 50 | 25 |
Table 4,2 (unit of culture medium：ng/ml)
| It is formulated E1 | It is formulated F1 | It is formulated G1 | It is formulated H1 | |
| Sall4B | 6 | 3 | 3 | 3 |
| SCF | 200 | 200 | 150 | 150 |
| FLT-3L | 300 | 200 | 150 | 100 |
| TPO | 50 | 25 | 25 | 12.5 |
Using the identical condition of culture of such as embodiment 1, application formula E/E1, is formulated F/F1 respectively, is formulated G/G1, is formulated H/
H1 cultivates human hematopoietic stem cell as culture medium.
As a result, CD34+ cells are averaged, amplification times seniority top digit is 65 times, and lower-order digit is 32 times；Therefore, expanding effect is more
It is preferable.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To be made various changes or modifications to the present invention, such equivalent forms equally fall within the model that the application the appended claims are limited
It encloses.
Claims (9)
1. a kind of kit for hematopoietic stem cell expansion culture, which is characterized in that including：Cell factor composition 1
With cell factor composition 2；The cell factor composition 1 is composed of the following components：
The cell factor composition 2 is composed of the following components：
2. kit as described in claim 1, which is characterized in that the cell factor composition 1 is composed of the following components：
3. kit as claimed in claim 2, which is characterized in that the cell factor composition 1 is composed of the following components：
4. kit as described in claim 1, which is characterized in that the cell factor composition 2 is composed of the following components：
5. kit as claimed in claim 4, which is characterized in that the cell factor composition 2 is composed of the following components：
6. the kit as described in Claims 1 to 5 is any, which is characterized in that further included in the kit：IMDM is cultivated
Base；The cell factor composition 1 and/or cell factor composition 2, which is divided in from IMDM culture mediums in different containers, to be put
It puts.
7. the kit as described in Claims 1 to 5 is any, which is characterized in that further included in the kit：IMDM is cultivated
Base；The cell factor composition 1 and/or cell factor composition 2 is formulated in IMDM culture mediums.
8. the kit as described in Claims 1 to 5 is any, which is characterized in that further include tire ox in the IMDM culture mediums
Serum.
9. the purposes of any kits of claim 1-8, which is characterized in that for hematopoietic stem cell expansion culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410066574.9A CN104862277B (en) | 2014-02-26 | 2014-02-26 | High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410066574.9A CN104862277B (en) | 2014-02-26 | 2014-02-26 | High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104862277A CN104862277A (en) | 2015-08-26 |
| CN104862277B true CN104862277B (en) | 2018-06-12 |
Family
ID=53908456
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410066574.9A Active CN104862277B (en) | 2014-02-26 | 2014-02-26 | High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104862277B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107663515B (en) * | 2016-07-28 | 2020-12-15 | 苏州方舟生物医药有限公司 | Method for directionally preparing human red blood cells and preparation |
| CN107384861B (en) * | 2017-08-31 | 2020-02-21 | 银丰生物工程集团有限公司 | Culture method for in vitro expansion of hematopoietic stem cells and enhancement of homing capacity of hematopoietic stem cells |
| CN108315300A (en) * | 2018-02-11 | 2018-07-24 | 大连金玛健康产业发展有限公司 | A kind of cell factor composition cultivates the application in candidate stem cell in vitro |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103124784A (en) * | 2010-08-23 | 2013-05-29 | 纽约州立大学研究基金会 | Method for expansion of stem cells and the use of such cells |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050032122A1 (en) * | 2003-08-06 | 2005-02-10 | Shiaw-Min Hwang | Optimizing culture medium for CD34<+> hematopoietic cell expansion |
-
2014
- 2014-02-26 CN CN201410066574.9A patent/CN104862277B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103124784A (en) * | 2010-08-23 | 2013-05-29 | 纽约州立大学研究基金会 | Method for expansion of stem cells and the use of such cells |
Non-Patent Citations (5)
| Title |
|---|
| Expansion of Cord Blood Stem Cells Mediated by Epigenetic Mechanisms Remain Permissive to External Environmental Cues;Hiroto Araki et al.;《BLOOD》;20081231;摘要 * |
| FLt-3 配基和促血小板生成素对脐血造血;刘英等;《中华儿科杂志》;20000228;第38卷(第2期);第89-91页 * |
| 人脐带血造血干细胞的分离和体外扩增;刘云霞等;《湖南师范大学学报(医学版)》;20041231;第1卷(第1期);摘要，表1-2 * |
| 扩增人脐血造血干/祖细胞重建SCID 小鼠造血的实验研究;刘英等;《中华血液学杂志》;19991231;第20卷(第12期);摘要，第634页右栏第2段至第635页左栏第1段 * |
| 造血细胞生长因子对脐血CD34+细胞短期体外扩增的作用;叶韵斌等;《福建医科大学学报》;20030630;第37卷(第2期);第147-150页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104862277A (en) | 2015-08-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101445337B1 (en) | Isolation and purification of hematopoietic stem cells from post-liposuction lipoaspirates | |
| CN101821383B (en) | Culture medium and method for in vitro culturing human adult primary mesenchymal stem cells on large scale, primary mesenchymal stem cells obtained by method, uses thereof | |
| CN105331575B (en) | A kind of efficient amplification cultivating system of human vascular endothelial progenitor cells | |
| CN109082411A (en) | A method of obtaining the macrophage with phagocytic function by pluripotent stem cell differentiation | |
| CN104862277B (en) | High-efficiency in-vitro human hematopoietic stem cell amplification cultivation formula of liquid | |
| CN102643784A (en) | Expansion system in vitro for hematopoietic stem/progenitor cell | |
| CN105713880A (en) | Serum-free culture medium for hematopoietic stem cell in vitro expansion culture and application thereof | |
| CN108504625A (en) | A kind of l cell and application thereof | |
| CN109370988A (en) | Ex vivo expansion of stem cell cultivating system and its method | |
| CN103509101B (en) | A kind of cytokine expanding umbilical cord blood hematopoietic stem cell and culture medium thereof | |
| CN110846277B (en) | Immortalized mouse microglial cell line B6Mi1 as well as establishment method and application thereof | |
| CN104862278B (en) | A kind of in vitro human hematopoietic stem cell amplification cultivation formula of liquid | |
| CN101684454A (en) | Method for preparing and separating definitive endoderm | |
| EP1743024B1 (en) | Method of expanding cord blood cells | |
| CN107663515B (en) | Method for directionally preparing human red blood cells and preparation | |
| CN104114694A (en) | Method for the ex vivo expansion of hematopoietic stem and progenitor cells | |
| CN110055214B (en) | Application of ginsenoside in promoting in-vitro proliferation of umbilical cord mesenchymal stem cells | |
| CN107201338B (en) | Method for inducing proliferation and erythroid differentiation of hematopoietic stem and progenitor cells and application thereof | |
| CN109652377B (en) | Preparation method and application of lung cancer stem cells | |
| WO2023143006A1 (en) | Kit for inducing ips cells into nk cells and use method thereof | |
| WO2020147272A1 (en) | Method for preparing heterogeneous hematopoietic stem/progenitor cells by non-mobilized peripheral blood | |
| CN109735488A (en) | A method of being quickly obtained the mescenchymal stem cell of a large amount of high-purities | |
| CN104498435B (en) | A kind of acute leaching of primary B cell is the extracorporeal culturing method of leukaemia cell | |
| CN112662616A (en) | Method for inducing and differentiating human mesenchymal stem cells into cartilage with high efficiency | |
| CN106282089B (en) | A kind of efficient amplification cultivating system of non-human primate endothelial progenitor cell |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| EXSB | Decision made by sipo to initiate substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190515 Address after: 215126 Qingqiu Street 277, Suzhou Industrial Park, Jiangsu Province Patentee after: Suzhou Ark Biomedicine Co., Ltd. Address before: 215126 Qingqiu Street 277, Suzhou Industrial Park, Jiangsu Province Patentee before: Suzhou Fangzhou Gene Pharmaceutical Co., Ltd. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201216 Address after: 215126 Qingqiu Street 277, Suzhou Industrial Park, Jiangsu Province Patentee after: Suzhou Ark Biotechnology Co.,Ltd. Address before: 215126 Qingqiu Street 277, Suzhou Industrial Park, Jiangsu Province Patentee before: Suzhou Ark Biomedicine Co.,Ltd. |
|
| TR01 | Transfer of patent right |