Cellular and Molecular Biology Core

CMBC Director: Nada Abumrad, PhD

CMBC Associate Director: Jun Yoshino, MD, PhD

CMBC Lab Manager: Terri Pietka, MA
Phone: 314-362-8459
Email: tpietka@wustl.edu

Location

The Cellular and Molecular Biology Core (CMBC) – formerly the Adipocyte Biology and Molecular Nutrition (ABMN) Core – occupies over 2000 square feet of space on the 2nd floor of the West Building within the main NORC space and the Division of Geriatrics and Nutritional Science. The CMBC can be reached by most NORC members by a short 5-10 minute walk through adjoining hallways and a 2nd floor pedestrian link that connects all research buildings on the Washington University School of Medicine Campus.

Visit the Cellular and Molecular Biology Core website


The purpose of the Cellular and Molecular Biology Core (CMBC) is to assist NORC investigators in evaluating the cellular and molecular mechanisms involved in: i) pathogenesis of obesity; ii) the multi-organ system complications associated with obesity; and iii) the multi-organ system abnormalities caused by alterations in nutrition and metabolism. One major focus of the services provided by the CMBC is cell-cell communication and tissue cross talk, because of the importance of inter-organ and cell-to-cell signaling in the pathogenesis of nutritional and obesity related diseases. The expertise of the CMBC directors in cellular metabolism, cell biology and molecular nutrition, the highly skilled staff, and the array of well-maintained specialized equipment provide NORC investigators with integration of the diverse services needed to study mechanisms in cell and tissue systems which will enhance both basic and clinical research studies.

The specific aims of the Cellular and Molecular Biology Core (CMBC) are to provide NORC investigators with the following services:

  • Cellular assessments needed for sophisticated nutrition/obesity research
    • Provision of cell lines (mouse and human pre-adipocytes, adipocytes, myoblasts, macrophages) with validated culture and differnetiation protocols
    • Evaluation of adipose tissue morphology
    • Isolation of adipocytes and stromal vascular fraction cells from adipose tissue explants/biopsies
    • Standardized preparation of tissue samples for gene expression and protein content; v) optimized protocols for metabolic studies in cells and tissue explants/biopsies (e.g. glucose and fatty acid uptake and release)
    • Training-assistance in cell/tissue respiration and mitochondrial function
    • Training-assistance in measurement of plasma adipokine, myokine, hepatokine and cytokine levels using multiplex technology
    • Training-assistance in isolation and characterization of extracellular vesicles (exosomes)
  • Access to specialized equipment
    • Much of the equipment offered by the CMBC for use by NORC members is too costly for individual investigators and not available elsewhere in the institution. CMBC staff provide NORC investigators and/or their technical staff with one-on-one training in equipment use, written standard operating procedures, and continued support in troubleshooting technical issues.
  • Expert consultative services to allow investigators at any level of experience to conduct sophisticated mechanistic nutrition/obesity research
    • The Core directors and staff have considerable expertise in molecular, cellular and integrative metabolism. Consultative services help basic and clinical investigators determine the most appropriate and robust experimental approaches and CMBC services needed to answer their nutrition/obesity related research questions. The consultative services also include assistance with troubleshooting and data interpretation.

Equipment

The major equipment items available through the Cellular and Molecular Biology Core (CMBC):

For expression analyses (gene and protein):

2      ABI QuantStudio real-time PCR

1      Li-Cor Odyssey Near IR protein detection system

1      Li-Cor C-DiGit Chemiluminescence protein detection system

1      Qiagen TissueLyser II

1      Nanodrop ND-1000 spectrophotometer

1      Bio-Tek Synergy II multi-mode plate reader

For adipocyte morphology, metabolic assays and cell respiration:

1      Aurora Scientific Single Fiber Analyzer

1      Beckman liquid scintillation counter

1      Beckman Coulter Multisizer III for adipocyte sizing

2      Oroboros Oxygraph 2K respirometry systems for mitochondrial physiology studies

For cell and organ communication:

1      Nanosight NS300 for extracellular vesicle analysis

1      Luminex 200 Multiplex Immunoassay Analyzer to measure cytokines, adipokines, etc., in  biospecimens

Assorted small equipment for sample processing (vortexers, heating blocks, water baths, analytical balances, stir plates, microfuges, etc.), centrifuges, refrigerators, liquid nitrogen cell freezers, and -80°C freezers, integrated into a remote monitoring and personnel notification system

Equipment in the CMBC tissue culture facility includes:

3      NuAire laminar flow hoods

4      NuAire CO2 incubators

1      Automated CO2 tank switcher

1      Lonza Amaxa Nucleofector II transfection system

1      World Precision Instruments EVOM-2 for cell monolayer electrical resistance measurements

1      Nikon TS100 light & fluorescent microscope

1      Life Technologies Countess automated cell counter 

Services available

The Cellular and Molecular Biology Core is committed to providing access to specialized equipment and technical services to facilitate nutrition and obesity research. If you have any questions regarding the CMBC for cost structure, please feel free to contact us.

Contact: Terri Pietka
Phone: 314-362-8459
Email: tpietka@wustl.edu 

Download CMBC Service Request form (pdf)

See a list of available services below.

1. Cells for Culture

  • 3T3-L1
  • 3T3-F442A
  • OP9
  • HIB1B
  • LS14
  • LiSa-2
  • SBGS
  • C2C12
  • HSMM

2. Adipose Tissue Morphology

3. Gene Expression Analysis and RNA Extraction

Pathways of Interest:

  • Autophagy
  • Fatty acid metabolism
  • Glucose metabolism
  • Endoplasmic reticulum stress
  • ECM/Fibrosis
  • Angiogenesis
  • Inflammation

4. Protein Expression Analyses and Lysate Preparation

Pathways of Interest:

  • mTOR signaling
  • Insulin signaling
  • Endoplasmic reticulum stress
  • Inflammation

5. Substrate Metabolic Assays

  • Glucose uptake
  • Glycolysis
  • Glucose oxidation
  • Glycogenesis
  • Fatty acid uptake
  • Fatty acid oxidation
  • Lipid incorporation
  • Lipolysis

6. Mitochondrial Physiology

Substrate/inhibitors:

  • Glutamate/Malate
  • Oligomycin
  • Pyruvate
  • Rotenone
  • Succinate
  • Antimycin A
  • Uncoupling

7. Microscopy/Cell imaging

8. Training

  • Biochemical characterization assays
  • Mitochondrial function and physiology assays
  • Cellular trafficking and imaging
  • Autophagy analysis

9. Project consultation

10. Equipment Usage

  • ABI-7500 Fast
  • LiCor Odyssey
  • Tissue Culture Facility
  • Nikon TE2000U Microscope
  • Spectroscopy (Bio-Tek/NanoDrop)
  • Oxygraph 2K
  • Qiagen TissueLyser II

 

Please acknowledge the Washington University NORC, Nutrition Obesity Research Center,  in your publications by: “Supported by NIH grant P30 DK056341 (Nutrition Obesity Research Center).”