Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct
Obesity, defined as a body mass index of 30 kg/m2 or above, has increased considerably in incidence and frequency within the United States and globally. Associated comorbidities including cardiovascular disease, type 2 diabetes mellitus, metabolic syndrome, and nonalcoholic fatty liver disease have...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2020-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2020/4242130 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849307561815179264 |
|---|---|
| author | Robert Bender Michelle McCarthy Theodore Brown Joanna Bukowska Stanley Smith Rosalyn D. Abbott David L. Kaplan Christopher Williams James W. Wade Andrea Alarcon Xiying Wu Frank Lau Jeffrey M. Gimble Trivia Frazier |
| author_facet | Robert Bender Michelle McCarthy Theodore Brown Joanna Bukowska Stanley Smith Rosalyn D. Abbott David L. Kaplan Christopher Williams James W. Wade Andrea Alarcon Xiying Wu Frank Lau Jeffrey M. Gimble Trivia Frazier |
| author_sort | Robert Bender |
| collection | DOAJ |
| description | Obesity, defined as a body mass index of 30 kg/m2 or above, has increased considerably in incidence and frequency within the United States and globally. Associated comorbidities including cardiovascular disease, type 2 diabetes mellitus, metabolic syndrome, and nonalcoholic fatty liver disease have led to a focus on the mechanisms promoting the prevention and treatment of obesity. Commonly utilized in vitro models employ human or mouse preadipocyte cell lines in a 2-dimensional (2D) format. Due to the structural, biochemical, and biological limitations of these models, increased attention has been placed on “organ on a chip” technologies for a 3-dimensional (3D) culture. Herein, we describe a method employing cryopreserved primary human stromal vascular fraction (SVF) cells and a human blood product-derived biological scaffold to create a 3D adipose depot in vitro. The “fat-on-chip” 3D cultures have been validated relative to 2D cultures based on proliferation, flow cytometry, adipogenic differentiation, confocal microscopy/immunofluorescence, and functional assays (adipokine secretion, glucose uptake, and lipolysis). Thus, the in vitro culture system demonstrates the critical characteristics required for a humanized 3D white adipose tissue (WAT) model. |
| format | Article |
| id | doaj-art-1ffd3751b9804188b44cbdcd9bf59e36 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-1ffd3751b9804188b44cbdcd9bf59e362025-08-20T03:54:43ZengWileyStem Cells International1687-966X1687-96782020-01-01202010.1155/2020/42421304242130Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat ConstructRobert Bender0Michelle McCarthy1Theodore Brown2Joanna Bukowska3Stanley Smith4Rosalyn D. Abbott5David L. Kaplan6Christopher Williams7James W. Wade8Andrea Alarcon9Xiying Wu10Frank Lau11Jeffrey M. Gimble12Trivia Frazier13LaCell LLC, New Orleans LA, USACenter for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans LA, USACenter for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans LA, USACenter for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans LA, USALaCell LLC, New Orleans LA, USADepartment of Biomedical Engineering, Carnegie Mellon University, Pittsburgh PA, USADepartments of Biomedical Engineering and Chemical and Biological Engineering, Bioengineering and Biotechnology Center, Tufts University, Medford MA, USADepartment of Pharmacology, Xavier University of Louisiana, New Orleans LA, USAPlastic and Reconstructive Surgery, Baton Rouge LA, USALaCell LLC, New Orleans LA, USALaCell LLC, New Orleans LA, USALSU Department of Plastic Surgery, New Orleans, LA, USALaCell LLC, New Orleans LA, USALaCell LLC, New Orleans LA, USAObesity, defined as a body mass index of 30 kg/m2 or above, has increased considerably in incidence and frequency within the United States and globally. Associated comorbidities including cardiovascular disease, type 2 diabetes mellitus, metabolic syndrome, and nonalcoholic fatty liver disease have led to a focus on the mechanisms promoting the prevention and treatment of obesity. Commonly utilized in vitro models employ human or mouse preadipocyte cell lines in a 2-dimensional (2D) format. Due to the structural, biochemical, and biological limitations of these models, increased attention has been placed on “organ on a chip” technologies for a 3-dimensional (3D) culture. Herein, we describe a method employing cryopreserved primary human stromal vascular fraction (SVF) cells and a human blood product-derived biological scaffold to create a 3D adipose depot in vitro. The “fat-on-chip” 3D cultures have been validated relative to 2D cultures based on proliferation, flow cytometry, adipogenic differentiation, confocal microscopy/immunofluorescence, and functional assays (adipokine secretion, glucose uptake, and lipolysis). Thus, the in vitro culture system demonstrates the critical characteristics required for a humanized 3D white adipose tissue (WAT) model.http://dx.doi.org/10.1155/2020/4242130 |
| spellingShingle | Robert Bender Michelle McCarthy Theodore Brown Joanna Bukowska Stanley Smith Rosalyn D. Abbott David L. Kaplan Christopher Williams James W. Wade Andrea Alarcon Xiying Wu Frank Lau Jeffrey M. Gimble Trivia Frazier Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct Stem Cells International |
| title | Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct |
| title_full | Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct |
| title_fullStr | Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct |
| title_full_unstemmed | Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct |
| title_short | Human Adipose Derived Cells in Two- and Three-Dimensional Cultures: Functional Validation of an In Vitro Fat Construct |
| title_sort | human adipose derived cells in two and three dimensional cultures functional validation of an in vitro fat construct |
| url | http://dx.doi.org/10.1155/2020/4242130 |
| work_keys_str_mv | AT robertbender humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT michellemccarthy humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT theodorebrown humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT joannabukowska humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT stanleysmith humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT rosalyndabbott humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT davidlkaplan humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT christopherwilliams humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT jameswwade humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT andreaalarcon humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT xiyingwu humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT franklau humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT jeffreymgimble humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct AT triviafrazier humanadiposederivedcellsintwoandthreedimensionalculturesfunctionalvalidationofaninvitrofatconstruct |