Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943
Summary: Human subcutaneous adipocytes are an attractive therapeutic target in regulating overall physiological homeostasis. However, the differentiation of primary human adipose-derived models remains challenging. Here, we present a protocol to differentiate primary subcutaneous adipose-derived pre...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672300271X |
| _version_ | 1797828100124311552 |
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| author | Ajit Regmi William Roell |
| author_facet | Ajit Regmi William Roell |
| author_sort | Ajit Regmi |
| collection | DOAJ |
| description | Summary: Human subcutaneous adipocytes are an attractive therapeutic target in regulating overall physiological homeostasis. However, the differentiation of primary human adipose-derived models remains challenging. Here, we present a protocol to differentiate primary subcutaneous adipose-derived preadipocytes from human subcutaneous adipocytes and to measure lipolytic activity. We describe steps for seeding of subcutaneous preadipocytes, removal of growth factors, induction and maturation of adipocytes, removal of serum/phenol red in media, and treatment of mature adipocytes. We then detail glycerol measurement in conditioned media and its interpolation.For complete details on the use and execution of this protocol, please refer to Coskun et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-09T12:59:38Z |
| format | Article |
| id | doaj.art-1a84975addb04659982b18b60a852554 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-09T12:59:38Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-1a84975addb04659982b18b60a8525542023-05-13T04:25:44ZengElsevierSTAR Protocols2666-16672023-06-0142102304Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943Ajit Regmi0William Roell1Insulin Discovery, Eli Lilly and Company, Indianapolis, IN 46225, USA; Corresponding authorInsulin Discovery, Eli Lilly and Company, Indianapolis, IN 46225, USA; Corresponding authorSummary: Human subcutaneous adipocytes are an attractive therapeutic target in regulating overall physiological homeostasis. However, the differentiation of primary human adipose-derived models remains challenging. Here, we present a protocol to differentiate primary subcutaneous adipose-derived preadipocytes from human subcutaneous adipocytes and to measure lipolytic activity. We describe steps for seeding of subcutaneous preadipocytes, removal of growth factors, induction and maturation of adipocytes, removal of serum/phenol red in media, and treatment of mature adipocytes. We then detail glycerol measurement in conditioned media and its interpolation.For complete details on the use and execution of this protocol, please refer to Coskun et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672300271XCell BiologyMetabolism |
| spellingShingle | Ajit Regmi William Roell Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 STAR Protocols Cell Biology Metabolism |
| title | Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 |
| title_full | Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 |
| title_fullStr | Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 |
| title_full_unstemmed | Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 |
| title_short | Differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by GIP or LY3437943 |
| title_sort | differentiation of human subcutaneous adipocytes and measurement of lipolytic function induced by gip or ly3437943 |
| topic | Cell Biology Metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S266616672300271X |
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