Primary Cultures of Skin, Liver, and Kidney Cells from White Sucker (Catostomus commersonii) and Creek Chub (Semotilus atromaculatus)
Presenter(s)
Morgan Dahlager
Abstract
The Catostomus commersonii, also known as the White Sucker, and the Semotilus atromaculatus, Creek Chub, are freshwater fish primarily located in the upper Midwest in the Great Lakes basin and Northeast United States. They are very commonly found in small ponds, streams, and lakes all over Minnesota often used as bait and have very similar physical characteristics. These fish are very useful by acting as bioindicators by determining the health of these ecosystems and are useful for screening for sensitivity to pollutants and pathogens. This procedure was done by removing three different organs from each species of which are liver, kidney, and skin and transferred to flasks containing Leibovitz's L-15 media with 20% FBS and a PenStrep, Gentamycin, and Nystatin combo to observe how they grow over 3 months. Harvesting these organs was done with assistance including usage of their lab and materials at the USFWS in Lacrosse, WI. Two attempts at these cell lines have been made. The first was not successful in producing a viable cell line of any of the three organs from six White Sucker fish. The second attempt, with many modifications to procedure, did end up successful with more viable cells. Twelve fish were used in total between the two species for the second attempt. Six fish were White Sucker and six Creek Chub. Identification of each species was by their body shape and color pattern. The second attempt had more success due to multiple changes in the protocol. Some of which include surface decontamination of the fish before organ removal, homogenizing organs instead of mincing using a razor blade, and refeeding with fresh media compared to older media. Over time, the skin cells proved to be the most efficient in both species, although Creek Chub cultures grew better than White Sucker, by growing homogenous monolayers that appeared healthy enough for subculturing using a 1:3 split ratio after two weeks once the confluency was up to 80% from a Creek Chub. White Sucker skin cells also grew homogenous monolayers but only got up to approximately 55% at week four. The liver cells also proved to be the next best cell line due to another monolayer being formed and a successful subculture attempt using a 1:2 split ratio after three weeks once the confluency was up to approximately 75% from a Creek Chub. The kidney cells were also successful in both species by growing multiple sections of cells in their flasks for observation but not enough for subculturing. The highest confluency of White Sucker kidney got to 20% by week four and Creek Chub at 10% by week four. One liver and one kidney flask became contaminated around week 2-3 after set up, both from the same White sucker, but changes to the surface decontamination protocol significantly reduced the rate of contamination from the first attempt which had two kidney flasks from White Suckers get contaminated by week two and five.
College
College of Science & Engineering
Department
Biology
Campus
Winona
First Advisor/Mentor
Casey Finnerty
Location
Kryzsko Great River Ballroom, Winona, Minnesota; United States
Start Date
4-23-2026 2:00 PM
End Date
4-23-2026 3:00 PM
Presentation Type
Poster Session
Format of Presentation or Performance
In-Person
Session
2b=2pm-3pm
Poster Number
12
Primary Cultures of Skin, Liver, and Kidney Cells from White Sucker (Catostomus commersonii) and Creek Chub (Semotilus atromaculatus)
Kryzsko Great River Ballroom, Winona, Minnesota; United States
The Catostomus commersonii, also known as the White Sucker, and the Semotilus atromaculatus, Creek Chub, are freshwater fish primarily located in the upper Midwest in the Great Lakes basin and Northeast United States. They are very commonly found in small ponds, streams, and lakes all over Minnesota often used as bait and have very similar physical characteristics. These fish are very useful by acting as bioindicators by determining the health of these ecosystems and are useful for screening for sensitivity to pollutants and pathogens. This procedure was done by removing three different organs from each species of which are liver, kidney, and skin and transferred to flasks containing Leibovitz's L-15 media with 20% FBS and a PenStrep, Gentamycin, and Nystatin combo to observe how they grow over 3 months. Harvesting these organs was done with assistance including usage of their lab and materials at the USFWS in Lacrosse, WI. Two attempts at these cell lines have been made. The first was not successful in producing a viable cell line of any of the three organs from six White Sucker fish. The second attempt, with many modifications to procedure, did end up successful with more viable cells. Twelve fish were used in total between the two species for the second attempt. Six fish were White Sucker and six Creek Chub. Identification of each species was by their body shape and color pattern. The second attempt had more success due to multiple changes in the protocol. Some of which include surface decontamination of the fish before organ removal, homogenizing organs instead of mincing using a razor blade, and refeeding with fresh media compared to older media. Over time, the skin cells proved to be the most efficient in both species, although Creek Chub cultures grew better than White Sucker, by growing homogenous monolayers that appeared healthy enough for subculturing using a 1:3 split ratio after two weeks once the confluency was up to 80% from a Creek Chub. White Sucker skin cells also grew homogenous monolayers but only got up to approximately 55% at week four. The liver cells also proved to be the next best cell line due to another monolayer being formed and a successful subculture attempt using a 1:2 split ratio after three weeks once the confluency was up to approximately 75% from a Creek Chub. The kidney cells were also successful in both species by growing multiple sections of cells in their flasks for observation but not enough for subculturing. The highest confluency of White Sucker kidney got to 20% by week four and Creek Chub at 10% by week four. One liver and one kidney flask became contaminated around week 2-3 after set up, both from the same White sucker, but changes to the surface decontamination protocol significantly reduced the rate of contamination from the first attempt which had two kidney flasks from White Suckers get contaminated by week two and five.
Comments
Dahlager, Morgan M