Isolation and Maintenance of Primate Embryonic Stem Cells

Marshall, Vivienne S.; Waknitz, Michelle; Thomson, James A.

doi:10.1385/1-59259-220-1:11

Cited by 7 publications

(7 citation statements)

References 7 publications

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“…The classic methods used for culturing human ES cells have not changed dramatically since these procedures were described for mouse and primate ES cells (Robertson, 1987; Marshall et al. 2001).…”

Section: Tissue Culture Of Embryonic Stem Cellsmentioning

confidence: 99%

“…Human ES cells are cultured on mitotically inactivated MEFs using medium supplemented with 20% fetal bovine serum (FBS). Although primate ES cells require much more meticulous care than mouse ES cells (Marshall et al. 2001), they can be cultured in large numbers, frozen and thawed, using 10% DMSO together with 20% FBS or 30% serum replacer, with reasonable survival rates, or frozen with vitrification, which permits full recovery of the frozen colonies (Reubinoff et al.…”

Section: Tissue Culture Of Embryonic Stem Cellsmentioning

confidence: 99%

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Derivation and spontaneous differentiation of human embryonic stem cells*

Amit¹,

Itskovitz‐Eldor²

2002

Journal of Anatomy

198

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Embryonic stem (ES) cells are unique cells derived from the inner cell mass of the mammalian blastocyst. These cells are immortal and pluripotent, retain their developmental potential after prolonged culture, and can be continuously cultured in an undifferentiated state. Many in vitro differentiation systems have been developed for mouse ES cells, including reproducible methods for mouse ES cell differentiation into haematopoietic and neural precursors, cardiomyocytes, insulin-secreting cells, endothelial cells and various other cell types. The derivation of new human ES cell lines provides the opportunity to develop unique models for developmental research and for cell therapies. In this review we consider the derivation and spontaneous differentiation of human ES cells.

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Section: Tissue Culture Of Embryonic Stem Cellsmentioning

confidence: 99%

Section: Tissue Culture Of Embryonic Stem Cellsmentioning

confidence: 99%

Derivation and spontaneous differentiation of human embryonic stem cells*

Amit¹,

Itskovitz‐Eldor²

2002

Journal of Anatomy

198

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“…Although more than 100 human ESC lines have been isolated and are either registered by NIH or in the scientific literature [4, , , –8], fewer than 25 NHP ESC lines have been derived. Among these are 16 that originated from blastocysts recovered by nonsurgical uterine flush procedures from rhesus monkeys ( Macaca mulatta ) [9, 10] or common marmosets ( Callithrix jacchus ) [11]. In the latter instance, the derivation of three additional ESC lines was reported recently [12].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Characterization of Novel Rhesus Monkey Embryonic Stem Cell Lines

et al. 2006

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ESCs are important as research subjects since the mechanisms underlying cellular differentiation, expansion, and self-renewal can be studied along with differentiated tissue development and regeneration in vitro. Furthermore, human ESCs hold promise for cell and tissue replacement approaches to treating human diseases. The rhesus monkey is a clinically relevant primate model that will likely be required to bring these clinical applications to fruition. Monkey ESCs share a number of properties with human ESCs, and their derivation and use are not affected by bioethical concerns. Here, we summarize our experience in the establishment of 18 ESC lines from rhesus monkey preimplantation embryos generated by the application of the assisted reproductive technologies. The newly derived monkey ESC lines were maintained in vitro without losing their chromosomal integrity, and they expressed markers previously reported present in human and monkey ESCs. We also describe initial efforts to compare the pluripotency of ESC lines by expression profiling, chimeric embryo formation, and in vitro-directed differentiation into endodermal, mesodermal, and ectodermal lineages.

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“…2), and the latter species have an advantage of their smaller size in handling and breeding. The methods of Thomson et al [21] for establishment of ES cell lines were almost the same as those used to derive mouse ES cell lines. For example, they used Such pluripotent cell lines can be maintained and grown for a long period in culture and induced to differentiate into various cell types, which can be used for cell transplantation therapy and other purposes in medical research.…”

Section: Mouse Monkey and Human Es Cellsmentioning

confidence: 99%

Embryonic Stem Cell Lines of Nonhuman Primates

Nakatsuji

Suemori

2002

The Scientific World JOURNAL

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Human embryonic stem (ES) cell lines have opened great potential and expectation for cell therapy and regenerative medicine. Monkey and human ES cell lines, which are very similar to each other, have been established from monkey blastocysts and surplus human blastocysts from fertility clinics.Nonhuman primate ES cell lines provide important research tools for basic and applicative research. Firstly, they provide wider aspects of investigation of the regulative mechanisms of stem cells and cell differentiation among primate species. Secondly, their usage does not need clearance or permission from the regulative rules in many countries that are associated with the ethical aspects of human ES cells, although human and nonhuman embryos and fetuses are very similar to each other. Lastly and most importantly, they are indispensable for animal models of cell therapy to test effectiveness, safety, and immunological reaction of the allogenic transplantation in a setting similar to the treatment of human diseases.So far, ES cell lines have been established from rhesus monkey (Macaca mulatta), common marmoset (Callithrix jacchus), and cynomolgus monkey (Macaca fascicularis), using blastocysts produced naturally or by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). These cell lines seem to have very similar characteristics. They express alkaline phosphatase activity and stage-specific embryonic antigen (SSEA)-4 and, in most cases, SSEA-3. Their pluripotency was confirmed by the formation of embryoid bodies and differentiation into various cell types in culture and also by the formation of teratomas that contained many types of differentiated tissues including derivatives of three germ layers after transplantation into the severe combined immunodeficiency (SCID) mice.The noneffectiveness of the leukemia inhibitory factor (LIF) signal makes culture of primate and human ES cell lines prone to undergo spontaneous differentiation and thus it is difficult to maintain these stem cell colonies. Also, these ES cells are more susceptible to various stresses, causing difficulty with subculturing using enzymatic treatment and cloning from single cells. However, with various improvements in culture methods, it is now possible to maintain stable colonies of monkey ES cells using a serum-free medium and subculturing with trypsin treatment. Under such conditions, cynomolgus monkey ES cell lines can be maintained in an undifferentiated state with a normal karyotype and pluripotency even after prolonged periods of culture over 1 year. Such progress should facilitate many aspects of stem cell research using both nonhuman primate and human ES cell lines.

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Isolation and Maintenance of Primate Embryonic Stem Cells

Cited by 7 publications

References 7 publications

Derivation and spontaneous differentiation of human embryonic stem cells*

Derivation and spontaneous differentiation of human embryonic stem cells*

Isolation and Characterization of Novel Rhesus Monkey Embryonic Stem Cell Lines

Embryonic Stem Cell Lines of Nonhuman Primates

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