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Pt 5020

Manufactured by Lonza
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Sourced in United States

The PT-5020 is a piece of laboratory equipment designed for temperature-controlled mixing and incubation. It features a stainless steel platform and a microprocessor-controlled heating and cooling system to maintain precise temperature settings. The PT-5020 can accommodate a variety of sample containers and is suitable for various applications in life science research and analysis.

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Lab products found in correlation

C57bl 6j mouse line, by Jackson ImmunoResearch (2 mentions) Sihnoct, by Thermo Fisher Scientific (2 mentions) Sicontrl, by Santa Cruz Biotechnology (2 mentions) Pt 8202, by Lonza (2 mentions) Pt 9502, by Lonza (2 mentions) Cl 173, by American Type Culture Collection (1 mentions) Pt 5005, by Lonza (1 mentions) Tib 67, by American Type Culture Collection (1 mentions) Mycoalert, by Lonza (1 mentions) Pt 2501, by Lonza (1 mentions) Cc 2511, by Lonza (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Nb1rgb, by RIKEN BioResource Center (1 mentions) Pt 8002, by Lonza (1 mentions)

7 protocols using pt 5020

1

Isolation and Characterization of Murine and Human Adipocyte Progenitors

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The C57BL/6J mouse line was obtained from Jackson Laboratories. All animal procedures were performed under the guidance and approval of the University of California (San Francisco) Animal Care and Use Committee. Age and sex are reported in the main text as well as discussed in the limitations of the study section. Mice were maintained in a UCSF vivarium with 12 h light/dark cycles and fed standard chow diets.
Cells: De-identified human subcutaneous preadipocytes were purchased (PT-5020, Lonza). Primary SVF and adipocyte progenitor cells were isolated from male mice as described in STAR Methods below. For adipogenesis assays, cells were cultured in DMEM with 10% FBS in incubators at 37°C with 5% CO2.
Chen M., Kim S., Li L., Chattopadhyay S., Rando T.A, & Feldman B.J. (2023). Identification of an adipose tissue-resident pro-preadipocyte population. Cell reports, 42(5), 112440.
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2

Isolation and Characterization of Murine and Human Adipocyte Progenitors

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The C57BL/6J mouse line was obtained from Jackson Laboratories. All animal procedures were performed under the guidance and approval of the University of California (San Francisco) Animal Care and Use Committee. Age and sex are reported in the main text as well as discussed in the limitations of the study section. Mice were maintained in a UCSF vivarium with 12 h light/dark cycles and fed standard chow diets.
Cells: De-identified human subcutaneous preadipocytes were purchased (PT-5020, Lonza). Primary SVF and adipocyte progenitor cells were isolated from male mice as described in STAR Methods below. For adipogenesis assays, cells were cultured in DMEM with 10% FBS in incubators at 37°C with 5% CO2.
Chen M., Kim S., Li L., Chattopadhyay S., Rando T.A, & Feldman B.J. (2023). Identification of an adipose tissue-resident pro-preadipocyte population. Cell reports, 42(5), 112440.
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3

Adipocyte Differentiation and Metabolic Assays

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Human subcutaneous preadipocytes were purchased (PT-5020, Lonza) and differentiated according to the manufacturer’s protocol. Briefly, confluent preadipocyte were treated with Preadipocyte Growth Medium-2 (PGM-2TM) (PT-8202, Lonza) containing 10% FBS, 2 mM glutamine, IBMX (1:1000, PT-9502, Lonza), indomethacin (1:500, PT-9502, Lonza), dexamethasone (1:1000, PT-9502 Lonza) and insulin (1:100, PT-9502, Lonza). Confluent cells in 48-well plate were transfected with 7.5 pM sihNOCT (Thermofisher, 1299001) or siContrl (Santa Cruz, sc-37007) for 24 h and then differentiated for 2 days before being used in Seahorse assays or lysed for RNA extraction and RT-qPCR analysis. A list of the primers used is in Table S1. For human cell Seahorse XF Cell mito stress test, cells were directly seeded into XFe24 cell culture microplates (V7).
Chen M., Kim S., Li L., Chattopadhyay S., Rando T.A, & Feldman B.J. (2023). Identification of an adipose tissue-resident pro-preadipocyte population. Cell reports, 42(5), 112440.
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4

Cellular Model Maintenance and Characterization

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All cellular models were incubated at 37 °C and maintained in an atmosphere containing 5% CO2 and in accordance with sterile cell culture practices. Cell lines were tested for Mycoplasma (MycoAlert™, cat# LT07-318, Lonza) every 2 weeks by using the manufacturer’s conditions. Cell lines were purchased from American Type Culture Collection (ATCC®). HEPG2 (ATCC, HB-8065, human liver carcinoma), BJ (ATCC, CRL-2522, normal human foreskin fibroblast), HEK-293 (ATCC, CRL-1573, transformed human embryonic kidney cell line); and murine models 3T3-L1 (ATCC, CL-173), J774.1 (ATCC# TIB-67) and MEF (ATCC# CRL-2907) cells. Cells were grown in medium (DMEM, EMEM, IMDM or Basal medium) containing 10% fetal bovine serum (FBS, Hyclone), 2 mmol L-glutamine (GlutaMAX™), and 1% penicillin/streptomycin (for murine models) or gentamicin sulfate (50 μg/mL for human preadipocytes) to densities recommended by ATCC. The 3T3-L1 murine fibroblast is an accepted cellular adipogenesis model since it competently undergoes adipogenesis after treatment with induction medium and experiments were conducted with passages 1–16. For The human subcutaneous preadipocytes (Poietics™, PT-5020, Lonza) and visceral preadipocytes (PT-5005, Lonza) were cultured according to the manufacturer’s instructions up to passage 3 (Figure S12).
Ling T., Miller D.J., Lang W.H., Griffith E., Rodriguez-Cortes A., Ayachi I.E., Palacios G., Min J., Miranda-Carboni G., Lee R.E, & Rivas F. (2019). Mechanistic Insight on the Mode of Action of Colletoic Acid. Journal of medicinal chemistry, 62(15), 6925-6940.
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5

Fibroblast and Adipocyte Cell Culture

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NB1RGB, normal human neonatal skin fibroblast cells, and NHDF-Adult, normal human adult skin fibroblasts, were obtained from RIKEN Bioresource Center (Tsukuba, Japan) and Lonza (#CC-2511, Lonza, USA), respectively, and cultured in Minimum Essential Medium alpha (Wako, Japan) supplemented with 10% fetal bovine serum and penicillin/streptomycin (100 U/ml and 100 μg/ml; Invitrogen, USA) at 37°C in a 5% CO2. MSCs (#PT-2501, Lonza, USA) and pre-adipocytes (#PT-5020, Lonza, USA) were cultured and differentiated in accordance with manufacturer's instructions. See section ‘Adipogenic induction’ for details.
Tomaru Y., Hasegawa R., Suzuki T., Sato T., Kubosaki A., Suzuki M., Kawaji H., Forrest A.R., Hayashizaki Y., Shin J.W, & Suzuki H. (2014). A transient disruption of fibroblastic transcriptional regulatory network facilitates trans-differentiation. Nucleic Acids Research, 42(14), 8905-8913.
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6

Adipogenesis of Human Preadipocytes

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Primary human subcutaneous preadipocytes were purchased from Lonza (#PT-5020; Walkersville, MD). Human preadipocytes were treated with differentiation medium (#PT-8002; Lonza), and differentiated to mature adipocytes as described previously (29 (link)). Total RNA was extracted from preadipocytes immediately before and 8 days after induction of adipogenesis. We also evaluated CTGF gene expression reported in SVF cells and adipocytes isolated from subcutaneous adipose tissue samples in publicly available microarray datasets obtained from the Gene Expression Omnibus (GEO) database (GSE8995; https://www.ncbi.nlm.nih.gov/geo/) (30 (link)).
Yoshino J., Patterson B.W, & Klein S. (2019). Adipose tissue CTGF expression is associated with adiposity and insulin resistance in people. Obesity (Silver Spring, Md.), 27(6), 957-962.
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7

Adipocyte Differentiation and Metabolic Assays

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Human subcutaneous preadipocytes were purchased (PT-5020, Lonza) and differentiated according to the manufacturer’s protocol. Briefly, confluent preadipocyte were treated with Preadipocyte Growth Medium-2 (PGM-2TM) (PT-8202, Lonza) containing 10% FBS, 2 mM glutamine, IBMX (1:1000, PT-9502, Lonza), indomethacin (1:500, PT-9502, Lonza), dexamethasone (1:1000, PT-9502 Lonza) and insulin (1:100, PT-9502, Lonza). Confluent cells in 48-well plate were transfected with 7.5 pM sihNOCT (Thermofisher, 1299001) or siContrl (Santa Cruz, sc-37007) for 24 h and then differentiated for 2 days before being used in Seahorse assays or lysed for RNA extraction and RT-qPCR analysis. A list of the primers used is in Table S1. For human cell Seahorse XF Cell mito stress test, cells were directly seeded into XFe24 cell culture microplates (V7).
Chen M., Kim S., Li L., Chattopadhyay S., Rando T.A, & Feldman B.J. (2023). Identification of an adipose tissue-resident pro-preadipocyte population. Cell reports, 42(5), 112440.
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