Derivation of Human Embryonic Stem Cell Lines in Serum Replacement Medium Using Postnatal Human Fibroblasts as Feeder Cells

Inzunza, José; Gertow, Karin; Strömberg, Marie A.; Matilainen, Eija; Blennow, Elisabeth; Skottman, Heli; Wolbank, Susanne; Ährlund‐Richter, Lars; Hovatta, Outi

doi:10.1634/stemcells.2004-0201

Cited by 228 publications

(152 citation statements)

References 26 publications

Supporting

Mentioning

150

Contrasting

Unclassified

Order By: Relevance

“…Teratoma formation experiments were done as described before 47,48 by implantation of B10 6 cells beneath the testicular capsule of a young (7-week-old) severe combined immunodeficiency SCID mouse or by injecting the cells subcutaneously in female SCID mice. Three-four animals were used for each cell line.…”

Section: Methodsmentioning

confidence: 99%

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

et al. 2014

Self Cite

View full text Add to dashboard Cite

Lack of robust methods for establishment and expansion of pluripotent human embryonic stem (hES) cells still hampers development of cell therapy. Laminins (LN) are a family of highly cell-type specific basement membrane proteins important for cell adhesion, differentiation, migration and phenotype stability. Here we produce and isolate a human recombinant LN-521 isoform and develop a cell culture matrix containing LN-521 and E-cadherin, which both localize to stem cell niches in vivo. This matrix allows clonal derivation, clonal survival and long-term self-renewal of hES cells under completely chemically defined and xeno-free conditions without ROCK inhibitors. Neither LN-521 nor E-cadherin alone enable clonal survival of hES cells. The LN-521/E-cadherin matrix allows hES cell line derivation from blastocyst inner cell mass and single blastomere cells without a need to destroy the embryo. This method can facilitate the generation of hES cell lines for development of different cell types for regenerative medicine purposes.

show abstract

Section: Methodsmentioning

confidence: 99%

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…In fact, there was a first peak of generation of new hESC lines in 2004, contributed mostly from one group in the USA [12], one in Korea [13] and two in Sweden [14,15], which accounted for 16, 9 and 10 of the total of 38 new cell lines derived that year, respectively (Fig. 1).…”

Section: Gathering Information From Human Embryonic Stem Cell Registriesmentioning

confidence: 99%

A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells

Fraga¹,

Araújo²,

Stabellini

et al. 2011

Stem Cell Rev and Rep

View full text Add to dashboard Cite

Since the derivation of the first human embryonic stem cell (hESC) lines by Thomson and coworkers in 1998, more than 1,200 different hESC lines have been established worldwide. Nevertheless, there is still a recognized interest in the establishment of new lines of hESC, particularly from HLA types and ethnic groups currently underrepresented among the available lines. The methodology of hESC derivation has evolved significantly since 1998, when human LIF (hLIF) was used for maintenance of pluripotency. However, there are a number of different strategies for the several steps involved in establishing a new line of hESC. Here we make a survey of the most relevant parameters used between 1998 and 2010 for the derivation of the 375 hESC lines deposited in two international stem cell registries, and able to form teratomas in immunocompromised mice. Although we identify some trends in the methodology for establishing hESC lines, our data reveal a much greater heterogeneity of strategies than what is used for derivation of murine ESC lines, indicating that optimum conditions have not been consolidated yet, and thus, hESC establishment is still an evolving field of research.

show abstract

“…human cells have been reported to be supportive of hESC growth [16,19,[38][39][40][41][42][43][44][45]. Human feeders have the potential to be cGMP-compliant and have been used to derive multiple clinical-grade human ESC lines [46].…”

Section: Discussionmentioning

confidence: 99%

Efficient Derivation and Genetic Modifications of Human Pluripotent Stem Cells on Engineered Human Feeder Cell Lines

Zou

Chou

Dowey

et al. 2012

Stem Cells and Development

View full text Add to dashboard Cite

Derivation of pluripotent stem cells (iPSCs) induced from somatic cell types and the subsequent genetic modifications of disease-specific or patient-specific iPSCs are crucial steps in their applications for disease modeling as well as future cell and gene therapies. Conventional procedures of these processes require co-culture with primary mouse embryonic fibroblasts (MEFs) to support self-renewal and clonal growth of human iPSCs as well as embryonic stem cells (ESCs). However, the variability of MEF quality affects the efficiencies of all these steps. Furthermore, animal sourced feeders may hinder the clinical applications of human stem cells. In order to overcome these hurdles, we established immortalized human feeder cell lines by stably expressing human telomerase reverse transcriptase, Wnt3a, and drug resistance genes in adult mesenchymal stem cells. Here, we show that these immortalized human feeders support efficient derivation of virus-free, integration-free human iPSCs and long-term expansion of human iPSCs and ESCs. Moreover, the drug-resistance feature of these feeders also supports nonviral gene transfer and expression at a high efficiency, mediated by piggyBac DNA transposition. Importantly, these human feeders exhibit superior ability over MEFs in supporting homologous recombination-mediated gene targeting in human iPSCs, allowing us to efficiently target a transgene into the AAVS1 safe harbor locus in recently derived integration-free iPSCs. Our results have great implications in disease modeling and translational applications of human iPSCs, as these engineered human cell lines provide a more efficient tool for genetic modifications and a safer alternative for supporting self-renewal of human iPSCs and ESCs.

show abstract

Derivation of Human Embryonic Stem Cell Lines in Serum Replacement Medium Using Postnatal Human Fibroblasts as Feeder Cells

Abstract: 2005;23:544-549

Cited by 228 publications

References 26 publications

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment

A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells

Efficient Derivation and Genetic Modifications of Human Pluripotent Stem Cells on Engineered Human Feeder Cell Lines

Contact Info

Product

Resources

About