Establishment, characterization, and transfection potential of a new continuous fish cell line (CAM) derived from the muscle tissue of grass goldfish (Carassius auratus)

Li, Na; Guo, Lili; Guo, Huarong

doi:10.1007/s11626-021-00622-1

Cited by 18 publications

(19 citation statements)

References 56 publications

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“…The primary muscle cells were maintained in culture continuously by subculturing. To establish a cell line without genetic modifications, the cell culture proceeded through a crisis of spontaneous immortalization transformation, as previously described by Li et al 12 . Li et al characterized the spontaneous immortalization crisis by a large-scale apoptosis/senescence, resulting in sporadic clusters of quiescent cells that then grow out of quiescence to reach more confluent areas 12 .…”

Section: Resultsmentioning

confidence: 99%

“…To establish a cell line without genetic modifications, the cell culture proceeded through a crisis of spontaneous immortalization transformation, as previously described by Li et al 12 . Li et al characterized the spontaneous immortalization crisis by a large-scale apoptosis/senescence, resulting in sporadic clusters of quiescent cells that then grow out of quiescence to reach more confluent areas 12 . From passages 37–43, the mackerel cells underwent a similar crisis, marked by increased incidence of senescent cell morphology with clusters of viable cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Continuous fish muscle cell line with capacity for myogenic and adipogenic-like phenotypes

Saad

Yuen

Joyce

et al. 2023

Sci Rep

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Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel (Scomber scombrus) skeletal muscle cell line (“Mack” cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 h (± 19.1 SD). After a spontaneous immortalization crisis from passages 37–43, the cells proliferated at doubling times of 24.3 h (± 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 and myosin heavy chain immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O staining and quantification of neutral lipids. New qPCR primers (HPRT, PAX3B, MYOD1, MYOG, TNNT3A, and PPARG) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Continuous fish muscle cell line with capacity for myogenic and adipogenic-like phenotypes

Saad

Yuen

Joyce

et al. 2023

Sci Rep

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

“…To establish a cell line without genetic modifications, the cell culture proceeded through a crisis of spontaneous immortalization transformation, as previously described by Li et al 12 . Li et al characterized the spontaneous immortalization crisis by a large-scale apoptosis/senescence, resulting in sporadic clusters of quiescent cells that then grow out of quiescence to reach more confluent areas 12 . From passages 37 to 43, the mackerel cells underwent a similar crisis, marked by increased incidence of senescent cell morphology with clusters of viable cells (Fig.…”

Section: Establishment Of Continuous Cell Culture Through Spontaneous...mentioning

confidence: 99%

“…Nine fish cell lines, from Acipenser transmontanus (white sturgeon), Caranx melampygus (bluefin trevally), Lates calcarifer (barramundi), Danio rerio (zebrafish), Carassius auratus (grass goldfish), Channa striata (snakehead murrel), Labeo catla (Indian catla), and Wallago attu (helicopter catfish), have been isolated from fish muscle 9,10 ; however, none of the cell lines are characterized as muscle cells, as they lack myogenic potential. While fish muscle cells from species such as Danio rerio (zebrafish), Salmo salar (Atlantic salmon), Carassius auratus (grass goldfish), and Oncorhynchus mykiss (rainbow trout) have been isolated, their applicability to cellular aquaculture has been limited [11][12][13][14] . These limitations stem from some of these species not being commonly consumed (e.g., zebrafish, goldfish), the cell population not demonstrating muscle function (e.g., the aforementioned cell lines, goldfish), or their limited growth potential (salmon, trout) as the cells were not demonstrated in long-term culture.…”

Section: Introductionmentioning

confidence: 99%

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Continuous Fish Muscle Cell Line with Capacity for Myogenic and Adipogenic-like Phenotypes

Saad

Yuen

Joyce

et al. 2022

Preprint

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Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel (Scomber scombrus) skeletal muscle cell line (Mack cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 hr (+/- 19.1 SD). After a spontaneous immortalization crisis from passages 37-43, the cells proliferated at doubling times of 24.3 hr (+/- 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 (PAX7) and myosin heavy chain (MHC) immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O (ORO) staining and quantification of neutral lipids. New qPCR primers (HPRT, PAX3B, MYOD1, MYOG, TNNT3A, and PPARG) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.

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Sculpting the future of meat: Biomaterial approaches and structural engineering for large‐scale cell‐based production

Jin,

Bennie,

Ogilvie

et al. 2023

Sustainable Food Proteins

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Novel foods are an increasing reality (and necessity) for our global food system and need to be protein‐rich for good nutrition. Cell‐based meats use the edible biomass of in vitro cultured animal cells harvested from the muscle tissue of live animals, removing the need to raise and slaughter animals. It is a viable alternative that is environmentally friendly, lowers resource consumption, and reduces health risks associated with traditional livestock farming. The cell‐based meat industry has boomed over the last 5 years with dozens of start‐ups being founded and millions of dollars in capital raised. While cell‐based meat faces technical, socio‐political, and regulatory challenges, it is a nascent technology with key technical challenges such as cell line stability, non‐animal‐derived culture media development, and bioprocessing for commercial‐scale uses. Here, we review the current field of cell‐based meat production at a lab scale and assess the feasibility and scalability of commercial production and the challenges that these production methods face. Moreover, we discuss the advancements in technologies for large‐scale manufacturing of cell‐based meat covering aspects including optimized culture media formulations, edible scaffold designs, and bioreactors for high‐density cell culture to reduce production costs.

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Establishment, characterization, and transfection potential of a new continuous fish cell line (CAM) derived from the muscle tissue of grass goldfish (Carassius auratus)

Cited by 18 publications

References 56 publications

Continuous fish muscle cell line with capacity for myogenic and adipogenic-like phenotypes

Continuous fish muscle cell line with capacity for myogenic and adipogenic-like phenotypes

Continuous Fish Muscle Cell Line with Capacity for Myogenic and Adipogenic-like Phenotypes

Sculpting the future of meat: Biomaterial approaches and structural engineering for large‐scale cell‐based production

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