Hepatic Progenitor Cell Lines from Allyl Alcohol‐Treated Adult Rats Are Derived from γ‐Irradiated Mouse STO Cells

Zhang, Mingjun; Sell, Stewart; Leffert, Hyam L.

doi:10.1634/stemcells.21-4-449

Cited by 10 publications

(26 citation statements)

References 23 publications

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“…Rat 7777 (American Type Culture Collection [ATCC] CRL-1601) and mouse Hepa 1-6 (ATCC CRL-1830), Swiss NIH3T3, STO (ATCC CRL-1503), and STO-derived 3(8)21 and 3(8)21-enhanced green fluorescent protein (EGFP) [16] cells were cloned and cultured by standard procedures [16].…”

Section: Cell Lines and Cell Culturementioning

confidence: 99%

“…In each instance, to eliminate potential and unforeseen problems of cell contamination or mix up, chain of custody, subculture history, and passage logs were maintained, and routine measurements of mouse markers were made as described elsewhere [16]. The cells were harvested at passage 23 in Ca 2+ /Mg 2+ /HCO 3 --free Hanks' buffered saline supplemented with 0.05% trypsin and 0.53 mM EDTA; viability was >99% [16]. Ten million washed cells in 0.2-0.5 ml were injected intrasplenically [21] without immunosuppression.…”

Section: Xenotransplantationmentioning

confidence: 99%

“…DNA was isolated from cultured cells or liver samples [16] or from whole livers homogenized with a Brinkmann Polytron using DNeasy Tissue Kits (Qiagen, Valencia, CA). Polymerase chain reaction (PCR) products were purified using Qiagen MinElute PCR Purification Kits and characterized on agarose minigels [25] using 100-bp DNA ladders (Promega, Madison, WI).…”

Section: Polymerase Chain Reaction and Restriction Digest Analysesmentioning

confidence: 99%

“…Polymerase chain reaction (PCR) products were purified using Qiagen MinElute PCR Purification Kits and characterized on agarose minigels [25] using 100-bp DNA ladders (Promega, Madison, WI). Mouse and rat mitochondrial COX1 and ß-actin targets were amplified by standard procedures [16].…”

Section: Polymerase Chain Reaction and Restriction Digest Analysesmentioning

confidence: 99%

“…Recent transplantation studies with stromal stem cells and progenitor cells of bone marrow [10] and neural origin [11] and attenuated major histocompatibility complex (MHC) expression in these and similar cells [12][13][14] suggest this may be possible. In this report, evidence is presented that shows that embryonic mouse STO cell lines [15,16] behave like liver progenitor cells in nonimmunosuppressed rats. These findings are provocative for their biological implications to liver stem cell biology and gene therapy [17] and also because mouse STO cells have been, and continue to be, widely used as feeder layers to support culture and development of embryoid bodies [18], embryonic stem cells [19], and liver progenitor cells [13,20].…”

Section: Introductionmentioning

confidence: 99%

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Embryonic Mouse STO Cell–Derived Xenografts Express Hepatocytic Functions in the Livers of Nonimmunosuppressed Adult Rats

et al. 2005

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Cells derived from embryonic mouse STO cell lines differentiate into hepatocytes when transplanted into the livers of nonimmunosuppressed dipeptidylpeptidase IV (DPPIV)-negative F344 rats. Within 1 day after intrasplenic injection, donor cells moved rapidly into the liver and were found in intravascular and perivascular sites; by 1 month, they were intrasinusoidal and also integrated into hepatic plates with approximately 2% efficiency and formed conjoint bile canaliculi. Neither donor cell proliferation nor host inflammatory responses were observed during this time. Detection of intrahepatic mouse COX1 mitochondrial DNA and mouse albumin mRNA in recipient rats indicated survival and differentiation of donor cells for at least 3 months. Mouse COX1 targets were also detected intrahepatically 4-9 weeks after STO cell injection into nonimmunosuppressed wild-type rats. In contrast to STO-transplanted rats, mouse DNA or RNA was not detectable in untreated or mock-transplanted rats or in rats injected with donor cell DNA. In cultured STO donor cells, DPPIV and glucose-6-phosphatase activities were observed in small clusters; in contrast, mouse major histocompatibility complex class I H-2Kq , H-2D q , and H-2L q and class II I-A q markers were undetectable in vitro before or after interferon gamma treatment. Together with H-2K allele typing, which confirmed the Swiss mouse origin of the donor cells, these observations indicate that mouse-derived STO cell lines can differentiate along hepatocytic lineage and engraft into rat liver across major histocompatibility barriers.

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Section: Cell Lines and Cell Culturementioning

confidence: 99%

Section: Xenotransplantationmentioning

confidence: 99%

Section: Polymerase Chain Reaction and Restriction Digest Analysesmentioning

confidence: 99%

Section: Polymerase Chain Reaction and Restriction Digest Analysesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Embryonic Mouse STO Cell–Derived Xenografts Express Hepatocytic Functions in the Livers of Nonimmunosuppressed Adult Rats

et al. 2005

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Unexpected artifacts raise new caution in stem cell culture research

Leffert¹,

Sell²

2004

Hepatology

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With this letter, we wish to raise caution about interpretations of results owing to artifacts that may occur when cell lines are derived by culturing on cell-derived feeder layers. We recently reported the apparent derivation of rat liver progenitor cell lines by co-culture of liver cells enriched in periportal oval cells after allyl alcohol injury with ␥-irradiated mouse STO (SIM [Sandoz inbred Swiss mouse]) fibroblast feeder layers. 1 These reported lines are now undergoing intensive reinvestigation in our laboratories and the results indicate that the cells were actually derived from the STO feeder layers. In a series of first stage RT-PCR and PCR, aimed at extending the key original findings of constitutive and inducible differentiation, we were unable to detect rat genes or rat gene products in any of the 11 reported cell strains and lines, including the clonal "rat" line 3(8)#21, and its clonal derivative 3(8)#21-EGFP, a line transduced with enhanced green fluorescent protein. Instead, only mouse genes were detected in all instances. These results were obtained both with purified and cloned putative "rat liver progenitor" cells grown on or off STO feeder layers (␥-irradiated at UCSD), and with the original frozen "rat" cells obtained from untampered-with vials derived from very early passages at the Albany Medical College. These and other unexpected observations, which tend to eliminate cell fusion as the source of this artifact, are described in detail elsewhere. 2 Ongoing observations of hepatocytic functions in these lines suggest that some aspects of the original report may yet be correct. For example, the line best characterized, 3(8)#21-EGFP, expresses several hepatocyte properties in vitro and following transplantation (manuscript in preparation). In conclusion, this surprising and previously unreported type of cell culture artifact should be considered by all investigators of cultured stem cell systems which require co-culture with mitotically "inhibited" feeder layer cell lines.

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The importance of knowing your identity: Sources of confusion in stem cell biology†

Grompe

2004

Hepatology

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Hepatic Progenitor Cell Lines from Allyl Alcohol‐Treated Adult Rats Are Derived from γ‐Irradiated Mouse STO Cells

Cited by 10 publications

References 23 publications

Embryonic Mouse STO Cell–Derived Xenografts Express Hepatocytic Functions in the Livers of Nonimmunosuppressed Adult Rats

Embryonic Mouse STO Cell–Derived Xenografts Express Hepatocytic Functions in the Livers of Nonimmunosuppressed Adult Rats

Unexpected artifacts raise new caution in stem cell culture research

The importance of knowing your identity: Sources of confusion in stem cell biology†

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