WO2007103763A2 - Procedes de generation et d'utilisation de profils de cellules souches - Google Patents

Procedes de generation et d'utilisation de profils de cellules souches Download PDF

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Publication number
WO2007103763A2
WO2007103763A2 PCT/US2007/063123 US2007063123W WO2007103763A2 WO 2007103763 A2 WO2007103763 A2 WO 2007103763A2 US 2007063123 W US2007063123 W US 2007063123W WO 2007103763 A2 WO2007103763 A2 WO 2007103763A2
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Prior art keywords
stem
cells
cell
progenitor
sample
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PCT/US2007/063123
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English (en)
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WO2007103763A3 (fr
Inventor
Ami Treves
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Bio Regenerate, Inc.
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Publication of WO2007103763A2 publication Critical patent/WO2007103763A2/fr
Publication of WO2007103763A3 publication Critical patent/WO2007103763A3/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5073—Stem cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5082—Supracellular entities, e.g. tissue, organisms
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere

Definitions

  • Embryonic stem cells are derived from blastocysts and arise in a very early stage of embryonic development. Embryonic stem cells can be grown in large numbers in culture but are difficult to control developmentally. In addition, their use is accompanied by perceived and unresolved ethical problems.
  • Adult stem cells can be found in various tissues of the adult body. Each tissue and organ in the body originates from a small population of ASCs which is committed to differentiate into the various cell types that compose the tissue. ASCs are a likely source of continuous normal tissue replenishment, as well as, recovery in case of damage or disease, throughout the life of the organism.
  • stem and progenitor cells are found in many tissues and are thought to be responsible for normal development of the organism, replenishment of normal tissues, normal functions and physiology, various hormonal cycles, certain age related developments, development of certain diseases, normal and pathological immune responses, and recovery from diseases and injuries.
  • HSCs hematopoietic stem cells
  • hematopoietic stem cells Various types of hematopoietic and non-hematopoietic stem and progenitor cells have been found in the human blood. Endothelial progenitor cells, mesenchymal stem cells as well as fibrocytes that can mediate tissue repair have all been reported. These cells can also be found in bone marrow in the circulation and in other organs.
  • HSCs from either bone marrow, peripheral blood or cord blood are widely used for replacement of ablated bone marrow and for treatment of malignant and genetic diseases.
  • Bone marrow derived hematopoietic stem cells have also been reported to maintain pluripotent potential for non-hematopoietic tissues in addition to HSC. It has recently been found that the bone marrow contains more primitive stem cells than HSCs that can differentiate into other tissues and organs. Some of these ASCs belong to a well characterized subpopulation termed mesenchymal stem cells (MSCs) that can differentiate into bone, cartilage and heart muscle cells. Other pluripotent stem cells have also been detected in bone marrow. Bone marrow derived hematopoietic stem cells have also been reported to maintain pluripotent potential for non-hematopoietic tissues.
  • MSCs mesenchymal stem cells
  • Such mixed populations of cells have also been isolated from cord blood, adult peripheral blood, fat tissue and other organs. Under various conditions, those mixed populations can give rise to additional tissues such as blood vessels, bone, cartilage, muscle, liver, nerve cells as well as insulin secreting Langerhans cells.
  • ASCs [0005] Mixed populations of ASCs have also been identified in various other tissues. Actually, every tissue and organ in the body contains stem cells that participate in intrinsic regeneration and repair during growth, trauma and disease.
  • Monocyte and macrophage like cell populations that express pluripotent potential have also been observed in the circulation of healthy adults.
  • Peripheral blood endothelial progenitor cells that secrete angiogenic growth factor and form tube like structures have been reported to be derived from monocyte/macrophages.
  • Non- hematopoietic MSCs and pluripotent stem cells have been reported in non-mobilized human peripheral blood and such cells were also reported to reside in the monocytes/macrophage compartment.
  • stem cell populations from these various sources are being tested clinically for treatment of diseases such as ischemic heart, neural injuries, neuro-degenerative diseases, diabetes, as well as other diseases that do not currently have effective treatments.
  • diseases such as ischemic heart, neural injuries, neuro-degenerative diseases, diabetes, as well as other diseases that do not currently have effective treatments.
  • stem cell populations will be most effective therapeutically in particular situations. It is known that in some cases pure stem cell lines have not been effective at all.
  • a stem cell profile which could include an analysis of the quality and quantity of various populations of stem and progenitor cells, which reside at any given time in a tissue, could provide a useful tool for the clinical assessment of health conditions, for prognosis of various diseases, for tailoring individual therapies and following treatment efficacy as these uses become correlated with those stem cell profiles.
  • methods of determining stem cell profiles from tissues isolated from individuals are needed. Ideally the methods will provide for phenotypic and functional characterization of the various populations of stem and progenitor cells in tissue samples, such as blood samples. The methods should also provide information about the viability of the various stem and progenitor cell lineages found in a sample, hi addition, such profiles should also be associated with data corresponding to health conditions, disease prognosis, therapeutic treatment results and the like.
  • a simple method for phenotypical and functional characterization of various mixed populations of stem and progenitor cells from tissue samples is disclosed, hi one aspect, the method can be used to identify the status of these cells and their functions in their native host organisms.
  • the method can be applied to blood tissue and can include a step of enriching the three stem cell lineages that are commonly found in both the bone marrow, cord blood and peripheral blood: the hematopoietic, endothelial and mesenchymal stem/progenitor cells.
  • the variety and population of stem cells can be identified by surface marker analysis for several defined stem cell markers, by analysis of characteristic ex vivo self renewal potential, as well as by genetic analysis such as by determining gene expression profile. Such a marker analysis can be used to obtain a functional profile of stem/progenitor populations for each cell sample under various health or clinical situations.
  • a series of profiles can be generated for individuals and can be included in a database along with associated health characteristics such that the database can be used as a resource in the diagnosis of health conditions and as an aid in following patient responses to treatment, among other uses.
  • the data in the profiles can also be associated with stem/progenitor cell samples in a stem cell storage and banking setting and thereby add value to the stem cell bank.
  • a database of profiles for certain populations can serve for statistical, epidemiological and health evaluations.
  • the present disclosure covers methods for quantifying and reporting compositions of adult stem/progenitor cell populations isolated from specific tissues. [0012] The present application also is directed to the process of quantifying the stem and progenitor cell populations and reporting the quantities to the customer.
  • the tissue from which a stem cell sample is obtained can be blood, placenta, ascitic fluid, skin, kidney, liver, muscle, neural tissue or fat tissue, or their mixtures.
  • the tissue is unmobilized peripheral blood.
  • Each of the disclosed methods can be used to generate a revenue for a business, hi addition, access to the disclosed database or the information in the profiles could also be given for a fee in order to generate a business revenue.
  • the present invention includes the identification of mixed populations of stem and progenitor cells from individuals and preferably from various individual tissues of an individual.
  • Suitable body tissues include any tissues that harbor stem or progenitor cells.
  • Exemplary tissues include blood, cord blood, blood buffy coat, placenta, amniotic fluid, ascitic fluid, skin, kidney, liver, muscle, neural tissue, fat, tooth pulp, and the like.
  • the cell populations can include hematopoietic cells, or hematopoietic committed cell lineages including lymphoid cells, erythroid cells, myeloid cells, monocytic cells, megakaryocyte cells, and the like.
  • Cell populations can also include mesenchymal cells, stromal cells, fibroblasts, endothelial cells, hematopoietic cells, hematopoietic committed cell lineages and their mixtures, for example.
  • Example 1 Many identification and quantitation methods are known in the art and can be used, so long as they can be used to determine the number of isolated cell populations or mixed populations of cells.
  • Examples 1 and 2 set forth exemplary methods.
  • One exemplary method set forth in Example 1 involves obtaining a blood sample and, following hemolysis, subjecting the sample to analysis by fluorescent activated cell sorting (FACS) according to standard methods.
  • FACS fluorescent activated cell sorting
  • Cell markers for which FACS information can be obtained include CD34+, CD34+38-, CDl 05, CD90, VEGFR2, VWF, VE Cadherin, and CD31, for example.
  • Other tissue samples containing stem and progenitor cells could also be used, such as cord blood and others.
  • body fluid such as blood
  • solid tissues such as skin, liver, muscle or other tissue biopsies
  • mechanical and enzymatic methods can be used to disrupt the tissue to obtain suitable cell suspensions.
  • hematopoietic stem cells can be identified as cells that contain CD 34 but that lack CD 38 markers
  • mesenchymal stem cells can be identified as cells that contain CD 105 and CD90
  • endothelial progenitor cells can be identified as cells that contain VEGFR2, VWF, VE Cadherin and CD31, for example.
  • New markers can also be incorporated into this analysis as they are discovered.
  • classifications can change with time and all changes and marker identification sets are considered to be encompassed by the present profiling methods and the profiles themselves.
  • Example 2 An example of another type of analysis is set forth in Example 2.
  • stem and progenitor cells from tissue samples are enriched after the tissue sample is obtained.
  • the cells from the sample are distributed into suitable growth media for growing all stem and progenitor cells in the sample together.
  • the enriched culture is then divided and individual samples of cells are incubated in more specialized media for growing specific types of stem or progenitor cells.
  • the cells can be divided and grown in media suitable for selective growth of hematopoietic stem cells, mesenchymal stem cells, endothelial stem and progenitor cells, and hematopoietic and immune stem and progenitor cells, respectively, as described in more detail in Example 2.
  • FACS analysis can be used to determine the number of cells displaying particular markers at any growth stage from any of the cultures.
  • an analysis could include both a FACS analysis of cells derived directly from the tissue sample in addition to analysis of samples enriched for stem and progenitor cells from the tissue sample. Such an analysis is diagramed in Table 1 below.
  • the profiling method does not necessarily result in the destruction of the entire tissue sample.
  • the cell sample for the disclosed analyses can be obtained from a sample obtained for stem cell storage and can be used to provide additional information about the stored sample.
  • the tissue or stem/progenitor cells can be used for a medical treatment or cryo-preserved for future medical needs or used in research, as desired.
  • Samples suitable for storage or use include any of the cell populations described herein, including tissue samples, enriched samples, suspensions flow direct sample, and the like.
  • the invention includes a profile listing which can include the quantities of the cells in the various stem or progenitor cell populations.
  • This information can be included in one or more reports.
  • a report could include any or all of the following information: 1) names of each stem/progenitor cell that can be identified in the tissue, 2) the quantity of stem/progenitor cells identified in the tissue sample, 3) the percent of each type of cell identified in the sample unit, 4) the quantities of each type of cell in a healthy population, 5) the number of cells identified that carry specific cell markers and 6) identification or quantitation of certain functions that are characteristic of the stem and progenitor cell populations, such as colony formation.
  • This information can be included in one or more reports and may provide a representation of the person's stem cell profile from the tissue at the time the sample was obtained.
  • the report(s) can contain information about the quantity and viability of the cells from direct issue samples and from enriched populations, as described above. Reports can also include information about the tissue and individual from which it was extracted and characteristics, such as health or disease characteristics, of the individual and the tissue or other information as desired. An exemplary report is shown in Table 2 in Example 3 below.
  • the stem cell profiles can be used to evaluate health conditions of individuals.
  • a series of stem cell profiles from individuals can be included in a database along with information relating to the health condition of the individuals at the time of, preceding or after the cell sample is taken. Once a sufficient amount of such data is obtained and incorporated into a database the profiles can be associated with health conditions. Once such a database is generated data from a new profile can be used to predict or diagnose the health conditions in the individuals from which they are obtained.
  • Such a database could be used to predict certain diseases, for prognostic evaluation of diseases, for predicting individual therapeutic responsiveness to drugs, to follow patient responses in treatments or for other uses, as desired.
  • stem cell profiles can be used to assess the potential of a person's cells for stem/progenitor- based treatments for diseases that can be treated by tissue regeneration, by protein replacement, or by coagulation factors.
  • diseases include diseases associated with defective biological processes such as cardiac ischemia, osteoporosis, chronic wounds, diabetes, neural degenerative diseases, neural injuries, bone or cartilage injuries, ablated bone marrow, anemia, liver diseases, hair growth, teeth growth, retinal disease or injuries, ear diseases or injury, muscle degeneration or injury, plastic surgery.
  • the treatment methods can be applied to cosmetic therapies including, filling of skin wrinkles, supporting organs, supporting surgical procedures, treating burns, and treating wounds, for example.
  • Specific treatment methods can include autologous or allogeneic therapies.
  • the stem cell profile can be used to assess the potential of using the person's cells for preparing a therapeutic agent containing a product secreted from the aforementioned cell populations.
  • the method can be accomplished by preparing a cell population by any of the methods described previously and incubating the cells in culture media for a period of time sufficient to generate secreted products.
  • the secreted products can then be isolated from the culture media by known methods which one of skill in the art can appreciate will depend upon the nature of the product.
  • the stem cell profile can be used to asses the potential of using the person's cells for methods of using the disclosed cell populations in gene therapy. Such methods can be accomplished by preparing cell populations by methods as described above. The cells can then be transfected with a recombinant DNA or other methods of gene manipulations and the modified cells can be introduced into a patient in need thereof.
  • the disclosed methods can be used to generate revenues.
  • the method can utilize any or all of the above disclosed methods including any or all of the information gathering methods, the data gathered and the profiles themselves or any other disclosed product or service.
  • the information obtained and the profiles can be stored in a database together or separately.
  • a fee can be charged for carrying out any of the disclosed methods on behalf of a third party such as a patient, or for providing some or all of the gathered information to a researcher, healthcare worker, health or life insurance company, provider of medical services, government body or any other entity that is interested enough in the information to pay for it, such that a business can generate revenue.
  • the information can be sold for the purpose of research, for development of medical treatments and diagnostics or for any other desired use.
  • This example demonstrates a method for determining a profile of stem and progenitor cells using minimal manipulation of the cells from blood.
  • a sample of venous blood of about 10 to about 20 ml can be obtained aseptically in a heparinized tube.
  • the sample can be withdrawn from a unit of blood or obtained directly from an individual.
  • the sample is then analyzed using fluorescent activated cell sorting techniques
  • Cell markers for which FACS information can be obtained include CD34+, CD34+38-, CD105, CD90, VEGFR2, VWF, VE Cadherin, and CD31, CD3, CD4, CD8, CD14, CD45 for example
  • This example demonstrates a method for determining a profile of stem and progenitor cells from a blood sample.
  • a sample of venous blood of about 20 to about 50 ml can be obtained aseptically in a heparinized tube.
  • the sample can be obtained from a unit of blood that was withdrawn for storage or directly from an individual.
  • the blood sample was then subjected to ficoll paque density gradient separation, using standard methods, to isolate mononuclear cells (MNC).
  • MNC mononuclear cells
  • the enriched mononuclear cells are then divided into four aliquots.
  • One aliquot is plated for direct hematopoietic colony formation in semisolid media, supplemented with hematopoietic growth factors.
  • About 2 x 10 5 cells per ml, are plated in semi-solid medium (methyl cellulose) supplemented with 10%FCS and growth factors that stimulate hematopoietic cell growth including, for example SCF, G-CSF, GM-CSF, IL-3, IL-6, EPO and their combinations.
  • a colony count can then be determined after about two weeks.
  • different colonies, distinguished by morphological criteria as are known can be counted for various progenitor lineages responsive to specific growth factors or combinations of growth factors.
  • Culture conditions for growing colonies include for example, plating 1-lOxlO 6 mononuclear cells in tissue culture media such as RPMI, methyl cellulose, antibiotics, serum such as fetal calf serum, growth factors as mentioned above and incubation for example for 1- 2 weeks in a humidified 5%CO2 incubator.
  • tissue culture media such as RPMI, methyl cellulose, antibiotics, serum such as fetal calf serum, growth factors as mentioned above and incubation for example for 1- 2 weeks in a humidified 5%CO2 incubator.
  • One aliquot of the enriched mononuclear cell culture is plated in tissue culture media for enrichment of endothelial progenitor cells.
  • Enriched mononuclear cells can be plated at about 4 x 10 5 cells per 2 ml in fibronectin precoated culture plates.
  • the media used can be EBM2 supplemented with pre-selected FCS (20%) and growth factors including growth factors that stimulate endothelial progenitor cell growth, such as VEGF, bFGF, IGFl about 2-20 nanogram per ml each and their combinations.
  • VEGF vascular endothelial progenitor cell growth
  • the cells can be collected with trypsin-EDTA and plated at 2 x 10 5 cells per 2 ml. After an additional 6-8 days the cells can be collected and separated into two fractions, one fraction to be analyzed by FACS for endothelial cell markers such as VEGFR-2, VWF, VE-cadherin, CD31, one fraction to be analyzed using a colony formation assay for each growth factor or combination to be studied, in a known method for endothelial progenitor colony formation assay(CFU-E).
  • FACS endothelial cell markers
  • CD31 a colony formation assay for each growth factor or combination to be studied
  • One aliquot of the enriched mononuclear cells culture can be plated in tissue culture media for enrichment of mesenchymal stem cells.
  • Mononuclear cells can be plated at a density of about 4 x 10 5 cells per 2 ml in tissue culture plates in tissue culture media such as low glucose MEM supplemented with pre-selected FCS (10-20%) and growth factors that stimulate mesenchymal stem cells such as: , bFGF, M-CSF, IL6, LIF, about 20-50 nanogram per ml each, , After about 3-4 days, the nonadherent cells can be removed and the adherent cells further incubated in the same media. Media is replaced about every 3-4 days.
  • the cells are collected with trypsin-EDTA and plated at about 2 x 10 5 per 2 mis. After an additional 6-8 days the cells are collected and separated into two fractions which are analyzed by FACS analysis for markers such as CD 105 and CD90 and in a colony formation assay (CFU-M).
  • CFU-M colony formation assay
  • tissue culture media for enrichment of the hematopoietic and immune stem/progenitor cell lineages.
  • the enriched mononuclear cell culture can be plated at a density of 1 x 10 5 cells per 2ml in RPMI culture media supplemented with pre-selected FCS (10%) and growth factors combination such as: IL-6, SCF, Flt3, TPO, each factor about 50 nanogram per ml.
  • Fresh media can be added about every 3 to 4 days.
  • Example 3 The following example demonstrates the use of the results obtained in Examples 1 and 2 that can be organized into a concise report of a stem cell profile obtained from a tissue sample.
  • the procedure set forth in Example 1 can be used to obtain data which can be used to fill each of the three columns across the first line of Table 2 for data obtained from the tissue sample that was analyzed prior to ex vivo enrichment.
  • Example 2 The procedure set forth in Example 2 can be used to obtain data which can be used to fill each of the three columns across the second and third line of Table 2 for data obtained from the tissue sample that was analyzed after ex vivo enrichment.

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Abstract

La présente invention concerne un procédé de caractérisation fonctionnelle et phénotypique de diverses populations de cellules souche et progénitrices provenant d'échantillons cellulaires, tels que des échantillons de tissu. Dans un aspect, le procédé peut être utilisé pour identifier le statut de ces cellules et leurs fonctions dans des individus. Le procédé, une fois appliqué au tissu sanguin, peut comprendre une étape visant à enrichir des lignées de cellules souche qui se trouvent communément à la fois dans la moelle osseuse et dans le sang périphérique : les cellules souche/progénitrices hématopoïétiques, endothéliales et mésenchymateuses. La variété et la population de cellules souches peuvent être identifiées grâce à l'analyse de marqueurs cellulaires pour plusieurs marqueurs engagés dans la lignée et dans des cellules souches définies et à l'analyse de caractéristiques ex vivo présentant un potentiel d'auto-régénérescence. De telles techniques peuvent être utilisées en vue d'obtenir un profil fonctionnel de populations progénitrices/souche pour chaque échantillon cellulaire, et pour diverses conditions cliniques. Le profil peut également être associé et ajouter de la valeur au stockage et à la création de banques de cellules souches. Des profils et des profils consécutifs pour des individus peuvent être intégrés dans une base de données parallèlement aux caractéristiques de santé des individus. La base de données peut être utilisée pour le suivi du patient ou une référence scientifique. Des profils de bases de données pour certaines populations peuvent servir d'évaluations statistiques, épidémiologiques et sanitaires.
PCT/US2007/063123 2006-03-06 2007-03-02 Procedes de generation et d'utilisation de profils de cellules souches WO2007103763A2 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109897815A (zh) * 2019-02-20 2019-06-18 北京京蒙高科干细胞技术有限公司 一种无需包被的脂肪内皮祖细胞的高效分离和培养方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020142457A1 (en) * 1999-12-28 2002-10-03 Akihiro Umezawa Cell having the potentiality of differentiation into cardiomyocytes
US20050208025A1 (en) * 2002-04-16 2005-09-22 Fleming William H Enhancement of hematopoietic stem cell survival
US20050266556A1 (en) * 2004-02-09 2005-12-01 Yoder Mervin C Isolation, expansion and use of clonogenic endothelial progenitor cells

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020142457A1 (en) * 1999-12-28 2002-10-03 Akihiro Umezawa Cell having the potentiality of differentiation into cardiomyocytes
US20050208025A1 (en) * 2002-04-16 2005-09-22 Fleming William H Enhancement of hematopoietic stem cell survival
US20050266556A1 (en) * 2004-02-09 2005-12-01 Yoder Mervin C Isolation, expansion and use of clonogenic endothelial progenitor cells

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109897815A (zh) * 2019-02-20 2019-06-18 北京京蒙高科干细胞技术有限公司 一种无需包被的脂肪内皮祖细胞的高效分离和培养方法

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