CN1878860B - 分离和培养来自脐带血的间充质干细胞的方法 - Google Patents
分离和培养来自脐带血的间充质干细胞的方法 Download PDFInfo
- Publication number
- CN1878860B CN1878860B CN200480033035XA CN200480033035A CN1878860B CN 1878860 B CN1878860 B CN 1878860B CN 200480033035X A CN200480033035X A CN 200480033035XA CN 200480033035 A CN200480033035 A CN 200480033035A CN 1878860 B CN1878860 B CN 1878860B
- Authority
- CN
- China
- Prior art keywords
- cord blood
- cell
- stem cell
- csf
- mesenchymal stem
- 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.)
- Expired - Fee Related
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0662—Stem cells
- C12N5/0665—Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/22—Colony stimulating factors (G-CSF, GM-CSF)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2303—Interleukin-3 (IL-3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/23—Interleukins [IL]
- C12N2501/2306—Interleukin-6 (IL-6)
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Developmental Biology & Embryology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Hematology (AREA)
- Rheumatology (AREA)
- Immunology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及利用对细胞治疗而言最为理想的脐带血来分离和培养间充质干细胞的方法。该方法包括在体积为大于45ml/单位的脐带血中添加抗凝剂,所述脐带血是在分娩后24小时内获得的脐带血;以αMEM(α-极限必需培养基)稀释所得的抗凝剂和脐带血的混合物,随后通过离心收集单核细胞;以及将获得的单核细胞在αMEM中进行悬浮培养,培养基中含有干细胞因子、GM-CSF(粒细胞巨噬细胞集落刺激因子)、G-CSF(粒细胞集落刺激因子)、IL-3(白细胞介素-3)和IL-6(白细胞介素-6)。
Description
技术领域
本发明涉及利用对细胞治疗而言最为理想的脐带血来分离和培养间充质干细胞(mesenchymal stem cells)的方法。
背景技术
间充质干细胞是指能够分化为例如骨、软骨、脂肪、神经和肌肉等的原始细胞,已知骨髓中含有大量的间充质干细胞。实际上,当前由骨髓中分离间充质干细胞,然后用于特定目的的研究或广泛用于多种疾病的临床试验。
尽管可自骨髓中容易地获得间充质干细胞,但在获得骨髓方面存在困难,且目前还难以解决与将干细胞植入其他个体时所出现的免疫排斥反应有关的问题。
而相对于骨髓而言,脐带血比较容易获得,同时,如果获得了数量巨大的脐带血单位,则有可能使用与患者的组织相容性基因相同或最为相似的脐带血干细胞,并由此可能解决与免疫排斥有关的问题。不过,相对于骨髓而言,自脐带血获得间充质干细胞比较困难,且因此在研究和临床应用方面均存在困难。
通常,作为用于分离脐带血中所含的极少量间充质干细胞的方法,一直主要采用的方法包括用密度梯度离心方法分离白细胞层,然后进行细胞培养。不过,在密度梯度离心过程中很可能造成细胞丢失,因此使得很难培养脐带血中的那些极少量的间充质干细胞。
作为分离和培养间充质干细胞的常规方法,可参照美国专利5,197,985和5,486,359,其公开了通过分离并纯化的来自人类骨髓的间充质干细胞并培养这些细胞以增殖间充质干细胞的方法。也就是说,美国专利5,197,985涉及用于诱导来自人类骨髓的间充质干细胞分化为骨形成细胞的方法,其包括:通过将骨髓样品加到含有刺激间充质细胞生长但不刺激其分化的因子的培养基中,并在培养过程中仅容许间充质干细胞选择性粘附于一种基质表面,由此提供来自人类骨髓的分离的、纯化的、并培养扩增的间充质干细胞;将所述来自人类骨髓的分离的、纯化的、并培养扩增的间充质干细胞加到多孔载体上;并将含有培养扩增的来自人类骨髓的间充质干细胞的多孔载体植入含有使得人类间充质干细胞分化为骨细胞所必需的因子的环境中,其中所述多孔载体包含羟磷灰石和磷酸三钙(tricalcium phosphate),且其中所述培养基包括添加了胎牛血清(FBS)的BGJb培养基或F-12Nutrient Mixture。此外,美国专利5,486,359公开了能够分化为一种以上组织类型(例如骨、软骨、肌肉或骨髓基质)的细胞的分离的人类间充质干细胞,分离、纯化和培养扩增人类间充质干细胞的方法,以及此类细胞的表征和用途,特别是其研究试剂、诊断和治疗用途。在该专利中,间充质干细胞来自骨髓,并培养于含有胎牛血清的BGJb培养基中。
发明内容
技术问题
因此,本发明是针对培养间充质干细胞中的问题而做出的,本发明的一个目的在于提供一种用于分离和培养间充质干细胞的可重复性(reproducible)方法,其中可获得来自脐带血的间充质干细胞,而不会在密度梯度离心过程中造成细胞丢失。
技术方案
根据本发明的一个方面,通过提供一种用于自脐带血分离和培养间充质干细胞的方法而实现了上述和其他的目的,所述方法包括以下步骤:
在体积为大于45ml/单位的脐带血中添加抗凝剂,所述脐带血是在分娩后24小时内获得的纯脐带血;
以α-极限必需培养基(αMEM)稀释所得的抗凝剂和脐带血的混合物,随后通过离心收集单核细胞;以及
将获得的单核细胞在αMEM培养基中进行悬浮培养,培养基中含有干细胞因子、GM-CSF(粒细胞巨噬细胞集落刺激因子)、G-CSF(粒细胞集落刺激因子)、IL-3(白细胞介素-3)和IL-6(白细胞介素-6)。
本发明提供了用于在24小时内自脐带血单位中分离和培养间充质干细胞的可重复性方法,所述脐带血单位收集自分娩后产妇。也就是说,其目的在于通过找出自细胞数相对较少的脐带血中分离和培养间充质干细胞的最佳条件,从而有助于使用脐带血来治疗难治性疾病。
为实现这一目的,通过将脐带血以2倍体积的αMEM培养基进行稀释,继以进行离心以收集单核细胞层,然后在所述αMEM培养基中培养单核细胞,所述培养基中添加了细胞生长因子、干细胞因子、GM-CSF、G-CSF、IL-3和IL-6,由此能够保护原始间充质干细胞,并将细胞培养成功率由使用常规方法时的10%提高到大约90%,由此完成了本发明。
附图说明
根据下面的详细描述并结合附图,可更加清楚地了解本发明的上述的以及其他的目的、特征和其他优点。
在附图中,图1至图5所示的照片(X 100)分别是根据本发明的方法培养脐带血间充质干细胞系5、7、10、15和25天后的结果。
最佳实施方式
现结合以下实施例来更加详细地描述本发明的用于自脐带血中分离和培养间充质干细胞的方法。提供这些实施例的目的仅在于示例性地说明本发明,不能将其理解为是对本发明的范围和实质的限制。
实施例
实施例1:分离和培养间充质干细胞
为了自脐带血分离单核细胞,将脐带血以2倍体积的αMEM(α-极限必需培养基,Jeil Biotech Services,Korea)进行稀释,转移至50ml的Falcon管中,并以300xg在室温离心10min。收集分离的血膜层(buffycoat layer),再以2倍体积的αMEM进行稀释,覆盖在Ficoll-Hypaque上,并以300xg在室温离心30min。
Ficoll-Hypaque是Ficoll(蔗糖聚合物)和Hypaque(sodium ditrizoate)的聚合物,其被广泛用于自血液中分离单核细胞。Ficoll-Hypaque的比重为1.077g/ml,其比单核细胞重但比红细胞轻,因此可以根据单核细胞与红细胞之间的不同比重而将两者分离。也就是说,如果将血液置于Ficoll-Hypaque上并离心,单核细胞会聚集在Ficoll-Hypaque上。
将通过这种密度梯度离心方法所获得的单核细胞重新置于洗涤αMEM中,其中没有混合添加物,以200xg离心10min,随后弃掉αMEM并洗涤,得到沉积于Falcon管底部的细胞。再次向其中加入αMEM,200xg离心10min,随后弃掉αMEM并再洗涤一次,得到沉积于Falcon管底部的细胞。
随后,向含有抗生素(1000U/ml青霉素G,1000μg/ml硫酸链霉素,Gibco-BRL)和抗真菌剂(0.25μg/ml两性霉素B)和2mM的谷氨酰胺(Sigma)的αMEM培养基中加入20%胎牛血清(FBS,Jeil BiotechServices,Korea)和干细胞因子(50ng/ml)、GM-CSF(粒细胞巨噬细胞集落刺激因子:10ng/ml)、G-CSF(粒细胞集落刺激因子:10ng/ml)、IL-3(白细胞介素-3:10ng/ml)和IL-6(白细胞介素-6:10ng/ml)等细胞生长因子,并将细胞以1×106/cm2的浓度重悬。
自培养5天的细胞组中去除悬浮的细胞,获得粘附的细胞后,将其在含有20%胎牛血清和抗生素的αMEM培养基中培养25天,每隔2天更换全部培养基,不进行洗涤。
图1至图5所示的照片(X 100)分别是根据本发明的方法培养脐带血间充质干细胞系5、7、10、15和25天后所观察到的结果。可以看出,在培养分离自脐带血的单核细胞时,培养后5天,观察到粘附于瓶底并在瓶底生长的细胞,在第7天,细胞形成集落并生长成具有不同形状的细胞。培养后10天,单核细胞分化为纺锤状细胞,而在培养后25天,这些纺锤状细胞通过细胞分裂和扩增成为彻底的间充质干细胞,由此完成了培养。
实施例2:培养的间充质干细胞的细胞表面抗原的表征
为了确定上述分离和培养的纺锤状间充质干细胞的细胞表面抗原的特征,通过FACS对细胞表面抗原进行分析。结果显示于表1。通过将发光的免疫抗原指示剂附着在感兴趣的细胞表面,FACS(荧光激活的细胞分选;一种流式细胞仪)可分析感兴趣的细胞特征,或可根据所需的目的仅分离具有特定抗原指示剂的细胞。
表1
指示剂 | 反应 |
CD14 | - |
CD34 | - |
CD45 | - |
SH2 | + |
SH3 | + |
C29 | + |
CD44 | + |
CD90 | + |
CD166 | (+) |
表1证实,对于根据本发明所分离和培养的干细胞,CD34、CD45和CD14(造血干细胞的特征性指示剂)呈阴性反应,SH2、SH3、CD29和CD44(间充质干细胞的特征性指示剂)呈阳性反应,而CD166呈弱阳性反应。对这一事实的解释为,其说明根据本发明所分离并培养的细胞是间充质干细胞。
实施例3:比较间充质干细胞培养的成功率
分别根据常规的方法和本发明的方法培养50单位的脐带血,并比较两者的细胞培养的成功率。结果显示于下面的表2。
具体而言,在常规方法中,脐带血被用于干细胞培养,其中对所用的脐带血的量以及自分娩至收集的时间均没有设定标准。此外,常规方法的培养基中没有添加细胞生长因子,除此之外,采用的细胞的分离过程与本发明的方法相似。
表2
常规 | 本发明 | |
所得的间充质干细胞单位的数量 | 1 | 49 |
培养成功率(%) | 2 | 98 |
从以上表2可以看出,通过对间充质干细胞培养的成功率进行比较发现,常规方法的成功率为2%,而本发明具有98%的高成功率。
工业实用性
如上所述,根据本发明,可以自细胞数相对缺乏的脐带血中有效地分离并培养间充质干细胞,因此,可将原本无用而被废弃的脐带血作为细胞治疗的一种重要的途径,用于治疗各种难治性疾病。
尽管已经示例性地公开了本发明的优选的实施方式,但本领域普通技术人员应该理解,在不脱离如所附权利要求书所述的本发明的范围和实质的情况下,可以存在各种更改、添加和替换。
Claims (3)
1.用于自脐带血中分离和培养间充质干细胞的方法,所述方法包括以下步骤:
在体积为大于45ml/单位的脐带血中添加抗凝剂,所述脐带血是在分娩后24小时内获得的纯脐带血;
以α-极限必需培养基(αMEM)稀释所得的抗凝剂和脐带血的混合物,随后离心收集单核细胞;以及
将获得的单核细胞在αMEM培养基中进行悬浮培养,该培养基中加入干细胞因子、GM-CSF(粒细胞巨噬细胞集落刺激因子)、G-CSF(粒细胞集落刺激因子)、IL-3(白细胞介素-3)和IL-6(白细胞介素-6)。
2.权利要求1的方法,其中将所述脐带血以2倍体积的αMEM培养基进行稀释,覆盖在Ficoll-Hypaque上,并进行离心以收集单核细胞。
3.权利要求1的方法,其中所述用于培养单核细胞的αMEM培养基还加入抗生素、抗真菌剂、谷氨酰胺和胎牛血清。
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020030079362 | 2003-11-11 | ||
KR1020030079362A KR100560340B1 (ko) | 2003-11-11 | 2003-11-11 | 제대혈로부터 중간엽 줄기세포의 분리 및 배양 방법 |
KR10-2003-0079362 | 2003-11-11 | ||
PCT/KR2004/002715 WO2005045010A1 (en) | 2003-11-11 | 2004-10-25 | Method of isolating and culturing mesenchymal stem cell derived from umbilical cord blood |
Publications (2)
Publication Number | Publication Date |
---|---|
CN1878860A CN1878860A (zh) | 2006-12-13 |
CN1878860B true CN1878860B (zh) | 2012-05-30 |
Family
ID=36642516
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN200480033035XA Expired - Fee Related CN1878860B (zh) | 2003-11-11 | 2004-10-25 | 分离和培养来自脐带血的间充质干细胞的方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US7704739B2 (zh) |
EP (1) | EP1687414B1 (zh) |
KR (1) | KR100560340B1 (zh) |
CN (1) | CN1878860B (zh) |
AT (1) | ATE471368T1 (zh) |
DE (1) | DE602004027754D1 (zh) |
WO (1) | WO2005045010A1 (zh) |
Families Citing this family (39)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7311905B2 (en) * | 2002-02-13 | 2007-12-25 | Anthrogenesis Corporation | Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells |
US20080152629A1 (en) * | 2000-12-06 | 2008-06-26 | James Edinger | Placental stem cell populations |
WO2004069172A2 (en) | 2003-01-30 | 2004-08-19 | The Government of the United States of America as represented by the Department of Veterans Affairs | Multilineage-inducible cells and uses thereof |
CN100377165C (zh) * | 2003-04-24 | 2008-03-26 | 皇家飞利浦电子股份有限公司 | 无创式左心室的容积测定 |
KR100534215B1 (ko) * | 2003-11-11 | 2005-12-08 | (주)히스토스템 | 냉동 보관된 제대혈로부터 중간엽 줄기세포의 분리 및 배양 방법 |
KR100668371B1 (ko) * | 2004-12-30 | 2007-01-12 | 한훈 | 제대혈 유래 줄기세포를 이용한 대머리 치료 |
KR100679950B1 (ko) * | 2005-05-18 | 2007-02-08 | 한훈 | 제대혈 유래 줄기세포와 만니톨을 이용한근위축성측삭경화증 환자의 세포치료용 조성물 |
DE602005017812D1 (de) * | 2005-08-26 | 2009-12-31 | Seoul Nat Univ Ind Foundation | Multipotente blutstammzellen aus der nabelschnur und zellbehandlungsmittel damit zur behandlung der ischämischen krankheit |
NZ597304A (en) * | 2005-10-13 | 2013-06-28 | Anthrogenesis Corp | Immunomodulation using placental stem cells |
KR100697326B1 (ko) * | 2005-12-02 | 2007-03-20 | 재단법인서울대학교산학협력재단 | Oct4 발현능을 가지는 제대혈 유래 다분화능 성체줄기세포 및 그의 제조방법 |
EP1976977B1 (en) | 2005-12-29 | 2015-07-08 | Anthrogenesis Corporation | Placental stem cell populations |
KR100791487B1 (ko) * | 2006-05-29 | 2008-01-03 | 연세대학교 산학협력단 | 제대혈로부터 간엽줄기세포의 분리 및 배양 방법 |
KR100812544B1 (ko) * | 2006-10-19 | 2008-03-13 | 재단법인서울대학교산학협력재단 | 제대혈 유래 다분화능 줄기세포 및 이를 함유하는 허혈성괴사질환 치료용 세포치료제 |
US20100068194A1 (en) * | 2006-10-24 | 2010-03-18 | Dong-Ku Kim | Composition for in vivo transplantation for treatment of human cervical cancer comprising mononuclear cells derived from umbilical cord blood |
CN101657206B (zh) | 2007-02-12 | 2013-07-03 | 人类起源公司 | 利用胎盘干细胞治疗炎性疾病 |
KR100888924B1 (ko) * | 2007-06-04 | 2009-03-16 | 주식회사 제이비줄기세포연구소 | 인간 제대혈 유래 중간엽 줄기세포와 섬유아세포 분별방법 |
CN101878034B (zh) | 2007-09-28 | 2013-11-20 | 细胞基因细胞疗法公司 | 使用人胎盘灌洗液和人来自胎盘的中间体自然杀伤细胞的肿瘤抑制 |
KR101077042B1 (ko) * | 2008-03-28 | 2011-10-26 | 경희대학교 산학협력단 | 중배엽 줄기세포의 증식을 촉진하기 위한 cd45+ 세포 또는 cd45+ 세포 배양액의 용도 |
CA2734237C (en) * | 2008-08-20 | 2019-07-02 | Anthrogenesis Corporation | Treatment of stroke using isolated placental cells |
KR20200011604A (ko) | 2008-08-20 | 2020-02-03 | 안트로제네시스 코포레이션 | 개선된 세포 조성물 및 그의 제조 방법 |
AU2009283161A1 (en) | 2008-08-22 | 2010-02-25 | Anthrogenesis Corporation | Methods and compositions for treatment of bone defects with placental cell populations |
US8367409B2 (en) | 2008-11-19 | 2013-02-05 | Anthrogenesis Corporation | Amnion derived adherent cells |
CN102822330A (zh) | 2010-01-26 | 2012-12-12 | 人类起源公司 | 使用胎盘干细胞对骨相关癌的治疗 |
SI2556145T1 (sl) | 2010-04-07 | 2017-01-31 | Anthrogenesis Corporation | Angiogeneza z uporabo placentnih matičnih celic |
WO2011127113A1 (en) | 2010-04-08 | 2011-10-13 | Anthrogenesis Corporation | Treatment of sarcoidosis using placental stem cells |
WO2012009422A1 (en) | 2010-07-13 | 2012-01-19 | Anthrogenesis Corporation | Methods of generating natural killer cells |
CN102154200B (zh) * | 2010-12-13 | 2012-10-03 | 中国人民解放军第三○二医院 | 临床治疗用脐带间充质干细胞的制备和保存 |
US8969315B2 (en) | 2010-12-31 | 2015-03-03 | Anthrogenesis Corporation | Enhancement of placental stem cell potency using modulatory RNA molecules |
CN113559126A (zh) | 2011-06-01 | 2021-10-29 | 人类起源公司 | 利用胎盘干细胞治疗疼痛 |
CN102559590B (zh) * | 2012-01-16 | 2013-11-06 | 遵义医学院附属医院 | 用两种培养基序贯培养人脐带血间充质干细胞的方法 |
US8940294B2 (en) | 2012-03-02 | 2015-01-27 | Tissuetech, Inc. | Methods of isolating and culturing stem cells |
CN102876630B (zh) * | 2012-11-05 | 2015-02-18 | 东南大学 | 一种高效分离和扩增人脐带血间充质干细胞的方法 |
EP3622960A1 (en) | 2013-02-05 | 2020-03-18 | Celularity, Inc. | Natural killer cells from placenta |
CN103451151B (zh) * | 2013-06-19 | 2016-01-20 | 武汉道培胎盘干细胞生物技术有限公司 | 一种培养人脐带间充质干细胞的方法 |
CN103555663B (zh) * | 2013-11-19 | 2015-07-22 | 武汉道培胎盘干细胞生物技术有限公司 | 一种培养人羊膜间充质干细胞的方法 |
KR101642776B1 (ko) * | 2014-04-09 | 2016-07-29 | 대한민국 | 줄기세포를 포함하는 항생 조성물 |
RU2644650C2 (ru) | 2014-12-01 | 2018-02-13 | Общество с ограниченной ответственностью "Т-Хелпер Клеточные Технологии" | Материал стволовых клеток и способ его получения |
RU2708329C2 (ru) | 2016-05-31 | 2019-12-05 | Общество с ограниченной ответственностью "Т-Хелпер Клеточные Технологии" | Материал стволовых клеток, композиции и способы применения |
CN111643526A (zh) * | 2020-07-14 | 2020-09-11 | 贵州金元泰生物科技有限公司 | 一种用于延缓卵巢衰老及防治老年痴呆和抑郁的间充质干细胞制剂 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6335195B1 (en) * | 1997-01-28 | 2002-01-01 | Maret Corporation | Method for promoting hematopoietic and mesenchymal cell proliferation and differentiation |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5197985A (en) | 1990-11-16 | 1993-03-30 | Caplan Arnold I | Method for enhancing the implantation and differentiation of marrow-derived mesenchymal cells |
US5486359A (en) * | 1990-11-16 | 1996-01-23 | Osiris Therapeutics, Inc. | Human mesenchymal stem cells |
CN1564864A (zh) * | 2001-08-07 | 2005-01-12 | 麒麟麦酒株式会社 | 造血干细胞的制备方法 |
-
2003
- 2003-11-11 KR KR1020030079362A patent/KR100560340B1/ko active IP Right Grant
-
2004
- 2004-10-25 AT AT04793573T patent/ATE471368T1/de active
- 2004-10-25 US US10/579,070 patent/US7704739B2/en not_active Expired - Fee Related
- 2004-10-25 WO PCT/KR2004/002715 patent/WO2005045010A1/en active Application Filing
- 2004-10-25 EP EP04793573A patent/EP1687414B1/en not_active Not-in-force
- 2004-10-25 CN CN200480033035XA patent/CN1878860B/zh not_active Expired - Fee Related
- 2004-10-25 DE DE602004027754T patent/DE602004027754D1/de active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6335195B1 (en) * | 1997-01-28 | 2002-01-01 | Maret Corporation | Method for promoting hematopoietic and mesenchymal cell proliferation and differentiation |
Non-Patent Citations (2)
Title |
---|
Hou LL等.Induction of umbilical cord blood mesenchymal stem cells into neuron-like cells in vitro.《INTERNATIONAL JOURNAL OF HEMATOLOGY》.2003,第78卷(第3期),256-261. * |
侯玲玲等.人脐血间充质干细胞体外扩增和向神经元样细胞定向诱导分化的研究.《中华血液学杂志》.2002,第23卷(第8期),415-419. * |
Also Published As
Publication number | Publication date |
---|---|
CN1878860A (zh) | 2006-12-13 |
KR20050045325A (ko) | 2005-05-17 |
US20070092967A1 (en) | 2007-04-26 |
ATE471368T1 (de) | 2010-07-15 |
EP1687414B1 (en) | 2010-06-16 |
DE602004027754D1 (de) | 2010-07-29 |
WO2005045010A1 (en) | 2005-05-19 |
KR100560340B1 (ko) | 2006-03-14 |
EP1687414A4 (en) | 2007-09-12 |
EP1687414A1 (en) | 2006-08-09 |
US7704739B2 (en) | 2010-04-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN1878860B (zh) | 分离和培养来自脐带血的间充质干细胞的方法 | |
Nair et al. | Isolation and generation of human dendritic cells | |
US5965436A (en) | Method of isolating mesenchymal stem cells associated with isolated megakaryocytes by isolating megakaryocytes | |
CN103330720B (zh) | 一种混合干细胞注射剂及其制备方法 | |
CN1878861B (zh) | 分离和培养来自冻存脐带血的间充质干细胞的方法 | |
US7919316B2 (en) | Hematopoietic stem cell identification and isolation | |
WO1998020731A9 (en) | Msc-megakaryocyte precursor composition and method of isolating mscs associated with isolated megakaryocytes by isolating megakaryocytes | |
Briquet et al. | Prolonged ex vivo culture of human bone marrow mesenchymal stem cells influences their supportive activity toward NOD/SCID-repopulating cells and committed progenitor cells of B lymphoid and myeloid lineages | |
Nilsson et al. | Mast cells and basophils | |
CN104726401A (zh) | 一种利用胎盘间充质干细胞提高脐血间充质干细胞培养成功率的方法 | |
CN111909898B (zh) | 一种人牙髓干细胞的分离和扩增方法及其应用 | |
Lucas et al. | A linkage between dendritic cell and T-cell development in the mouse thymus: the capacity of sequential T-cell precursors to form dendritic cells in culture | |
CN110055219B (zh) | 一种利用非动员外周血制备异质性造血干祖细胞的方法 | |
CN104109653A (zh) | 利用无动物血清培养体系大规模扩增人外周血dnt细胞的方法 | |
CN103509753B (zh) | 一种人体造血干细胞分化培养方法 | |
CN102876632B (zh) | 人nk/t细胞系 | |
CN112300992B (zh) | 一种nk细胞培养液及多级激活nk细胞的培养方法 | |
KR100818215B1 (ko) | Cd34양성 줄기세포로부터 t 임파구 전구체를 제조하는 방법 | |
Tedder et al. | Isolation and generation of human dendritic cells | |
US4950598A (en) | Process for making T cell hybridomas | |
KR20220064163A (ko) | 세포 배양 면적 조절을 이용한 자연살해세포의 증식 촉진 방법 | |
CN117126809A (zh) | 体外诱导nk细胞扩增试剂盒及体外诱导nk细胞扩增方法 | |
Barkhatov et al. | Composition and functional properties of monolayer cell culture from human umbilical cord blood | |
OGAWA et al. | IN VITRO (EX VIVO) EXPANSION OF MURINE HEMATOPOIETIC STEM CELLS Running Title: In Vitro Expansion of Stem Cells | |
Chen | Analysis of the phenotype of elicited macrophages capable of in vitro proliferation |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120530 Termination date: 20141025 |
|
EXPY | Termination of patent right or utility model |