Immortalized embryonic hepatocytes were cultured in Medium 199 (WelGENE) that was supplemented with 10% fetal bovine serum (WelGENE) and 1% penicillin-streptomycin (WelGENE), as previously described (Back et al., 2009 (link)). AML12 muse normal hepatocytes were obtained from the American Type Culture Collection (ATCC, USA). The cells were cultured in DMEM/F12 (WelGENE) that was supplemented with 10% fetal bovine serum (WelGENE), 100 nM dexamethason (Sigma-Aldrich), 1% insulin-transferrin-selenium-pyruvate supplement (ITSP; WelGENE), and 1% penicillin-streptomycin (WelGENE). The Lenti-X 293T Cell Line (Clontech, USA) was cultured in DMEM (WelGENE) containing 10% fetal bovine serum (WelGENE) and 1% penicillin-streptomycin (WelGENE).
Insulin transferrin selenium pyruvate supplement itsp
Manufactured by Welgene
Sourced in Cameroon
The Insulin-Transferrin-Selenium-Pyruvate Supplement (ITSP) is a laboratory product designed to provide a combination of key components essential for cell culture applications. The supplement contains insulin, transferrin, selenium, and pyruvate, which collectively support various cellular processes and maintain a balanced growth environment for cultured cells.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640, by Welgene (2 mentions)
Lenti x 293t cell line, by Takara Bio (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
Dulbecco modified eagle medium (dmem), by Welgene (1 mentions)
Medium 199, by Welgene (1 mentions)
Fetal bovine serum (fbs), by Welgene (1 mentions)
Dexamethason, by Merck Group (1 mentions)
Dmem f12, by Welgene (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Oleate, by Merck Group (1 mentions)
Recombinant mouse interferon γ, by Merck Group (1 mentions)
Recombinant human klotho protein, by R&D Systems (1 mentions)
4 protocols using insulin transferrin selenium pyruvate supplement itsp
1
Immortalized Hepatocyte Culture Protocol
2
Culturing and Differentiating Immortalized Podocytes
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Dr. Peter Mundel kindly provided conditionally immortalized mouse podocytes [22 (link)]. We cultured the podocytes at 33°C under permissive conditions in DMEM containing 10% FBS and 10 U/mL of mouse recombinant interferon-γ (Sigma-Aldrich, St. Louis, MO, USA) to enhance the expression of a thermosensitive T antigen. For differentiation, we cultured podocytes under nonpermissive conditions at 37°C without interferon-γ for 14 days. We maintained these cells under serum-deprived conditions for 24 hours, treated them with 400 μM of palmitate (Pal) with or without 400 pM of recombinant human klotho protein (R&D systems, USA), oleate (Sigma-Aldrich, St. Louis, MO, USA), or NAC (Sigma-Aldrich) for 24 hours. We then harvested them for the next assay.
Dr. Moin A. Saleem (University of Bristol, Bristol, UK) generously provided human conditionally immortalized podocytes (AB8/23) [23 (link)]. We cultivated the human podocytes at 33°C (permissive conditions) in an RPMI-1640 medium supplemented with 10% FBS and Insulin-Transferrin-Selenium-Pyruvate Supplement (ITSP; WelGENE Inc., Daegu, South Korea) to induce expression of a thermosensitive T antigen. For differentiation, we maintained podocytes at 37°C (non-permissive conditions) without ITSP for 14 days. We grew immortalized human tubule cells in a DMEM/F12 medium containing 10% FBS.
Dr. Moin A. Saleem (University of Bristol, Bristol, UK) generously provided human conditionally immortalized podocytes (AB8/23) [23 (link)]. We cultivated the human podocytes at 33°C (permissive conditions) in an RPMI-1640 medium supplemented with 10% FBS and Insulin-Transferrin-Selenium-Pyruvate Supplement (ITSP; WelGENE Inc., Daegu, South Korea) to induce expression of a thermosensitive T antigen. For differentiation, we maintained podocytes at 37°C (non-permissive conditions) without ITSP for 14 days. We grew immortalized human tubule cells in a DMEM/F12 medium containing 10% FBS.
3
Immortalized Human Podocyte Culture Protocol
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Human conditionally immortalized podocytes (AB8/23), primarily cloned from human glomerular cultures, were characterized and generously provided by Dr. Moin A. Saleem (University of Bristol, Bristol, UK). Human podocytes were maintained in RPMI 1640 (WelGENE Inc., Daegu, South Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS), Insulin-Transferrin-Selenium-Pyruvate Supplement (ITSP; WelGENE Inc.), and antibiotics. Fresh media was supplied once every 2 days.
To stimulate human podocyte proliferation, cells were cultivated at 33℃ (permissive conditions) in a culture medium supplemented with human recombinant ITSP to induce expression of temperature-sensitive large T antigens. To induce differentiation, podocytes were maintained at 37℃ (non-permissive conditions) for at least 2 weeks, and for subcultures, 0.05% trypsin was used to detach cells from the culture dishes.18 (link)
To stimulate human podocyte proliferation, cells were cultivated at 33℃ (permissive conditions) in a culture medium supplemented with human recombinant ITSP to induce expression of temperature-sensitive large T antigens. To induce differentiation, podocytes were maintained at 37℃ (non-permissive conditions) for at least 2 weeks, and for subcultures, 0.05% trypsin was used to detach cells from the culture dishes.18 (link)
4
Cultivation and Differentiation of Human Podocytes
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Human conditionally immortalized podocytes (AB8/23), primarily cloned from human glomerular cultures, were characterized and generously provided by Dr. Moin A. Saleem (University of Bristol, Bristol, UK). Human podocytes were maintained in RPMI 1640 (WelGENE, Daegu, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS), Insulin-Transferrin-Selenium-Pyruvate Supplement (ITSP; WelGENE), and antibiotics. Fresh media was supplied once every two days.
To stimulate human podocyte proliferation, cells were cultivated at 33℃ (permissive conditions) in a culture medium supplemented with human recombinant ITSP to induce expression of temperature-sensitive large T antigens. To induce differentiation, podocytes were maintained at 37℃ without ITPS (non-permissive conditions) for at least two weeks and for subculture, 0.05% trypsin was used to detach cells from the culture dishes [12] .
To stimulate human podocyte proliferation, cells were cultivated at 33℃ (permissive conditions) in a culture medium supplemented with human recombinant ITSP to induce expression of temperature-sensitive large T antigens. To induce differentiation, podocytes were maintained at 37℃ without ITPS (non-permissive conditions) for at least two weeks and for subculture, 0.05% trypsin was used to detach cells from the culture dishes [12] .
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