Establishment and characterization of fibroblast cell line of the Songliao Black pig

The Songliao Black pig was the first lean-meat maternal breed bred in China. To preserve genetic diversity of this breed, we designed a procedure to use ear marginal tissue of one male pig to establish a fibroblast cell line (designated SBPF18), which was subsequently characterized according to the population doubling time, karyotype analysis, isoenzyme assay, DNA fingerprint and sex determination, meanwhile microbial contamination assays were conducted to demonstrate the cell line was contamination-free. This method is suitable for establishing the fibroblast cell line from small amounts of tissue, and could further facilitate genomic studies and genetic modification.Key words: Songliao Black pig, fibroblast cell line, primary explants technique, cryogenic preservation

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Establishment and characterization of a fibroblast cell line derived from Texel sheep

Biochemistry and Cell Biology ◽

10.1139/o09-005 ◽

2009 ◽

Vol 87 (3) ◽

pp. 485-492 ◽

Cited By ~ 11

Author(s):

Linfeng F. Li ◽

Weijun J. Guan ◽

Han Li ◽

Xueyan Z. Zhou ◽

Xiujuan J. Bai ◽

...

Keyword(s):

Cell Line ◽

Fluorescent Proteins ◽

Transfection Efficiency ◽

Chromosome Analysis ◽

Fibroblast Cell ◽

Population Doubling ◽

Established Cell Line ◽

Population Doubling Time ◽

Fibroblast Cell Line ◽

Plasmid Transfection

A Texel sheep ear marginal tissue fibroblast cell line (named TSF19) was successfully established by using a primary explant technique and cell cryoconservation technology. TSF19 cells were adherent, with a population doubling time of 24.9 h. Chromosome analysis showed that >90% of cells were diploid prior to cell passage 4. Isoenzyme analyses of lactate dehydrogenase and malate dehydrogenase showed that the TSF19 cells had no cross-contamination with other species. Tests for cell line contamination with bacteria, fungi, or mycoplasmas were also negative. Plasmids encoding the fluorescent proteins pEGFP-N3, pECFP-N1, pDsRed1-N1, and pEYFP-N1 were transfected into cells to study exogenous gene expression in the cells. The plasmid transfection efficiency was between 21.8% and 46.5%. This newly established cell line will not only preserve the genetic resources of the important Texel sheep at the cell level but will also provide a valuable resource for genomic, postgenomic, somatic cloning research.

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Establishment and characterization of fibroblast cultures derived from a female common hippopotamus (Hippopotamus amphibius) skin biopsy

10.1101/2020.10.14.338632 ◽

2020 ◽

Author(s):

Tao Wang ◽

Zelong Li ◽

Jinpu Wei ◽

Dongmin Zheng ◽

Chen Wang ◽

...

Keyword(s):

Cell Cultures ◽

Cultured Cells ◽

Population Decline ◽

Fibroblast Cell ◽

Population Doubling ◽

Population Doubling Time ◽

Fibroblast Cultures ◽

Hippopotamus Amphibius ◽

The Common

AbstractThe population decline in the common hippopotamus (Hippopotamus amphibius) has necessitated the preservation of their genetic resources for species conservation and research. Of all actions, cryopreservation of fibroblast cell cultures derived from animal biopsy is considered a simple but efficient means. Nevertheless, preserving viable cell cultures of the common hippopotamus has not been achieved to our knowledge. To this end, we detailed a method to establish fibroblast cell cultures from a female common hippopotamus fetus in this study. By combining the classic tissue explant direct culture and enzymatic digestion methods, we isolated a great number of cells with typical fibroblastic morphology and high viability. Characterization of the fibroblast cultures was carried out using different techniques. In short, neither bacteria/fungi nor mycoplasma was detectable in the cell cultures throughout the study. The population doubling time was 23.9 h according to the growth curve. Karyotyping based on Giemsa staining showed that cultured cells were diploid with 36 chromosomes in all, one pair of which was sex chromosomes. Mitochondrial cytochrome C oxidase subunit I gene sequence of the cultured cells was 99.26% identical with the Hippopotamus amphibius complete mitochondrial DNA sequence registered in GenBank, confirming the cells were derived from a common hippopotamus. Flow cytometry and immunofluorescence staining results revealed that the detected cells were positive for fibroblast markers, S100A4 and Vimentin. In conclusion, we isolated and characterized a new fibroblast cell culture from a common hippopotamus skin sample and the cryopreserved cells could be useful genetic materials for the future research.

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