Home › Cell Biology › Isolation and Culture of Human Mature Adipocytes Using Membrane Mature Adipocyte Aggregate Cultures (MAAC)
Cell Biology JoVE (Open Access) Citable · DOI

Isolation and Culture of Human Mature Adipocytes Using Membrane Mature Adipocyte Aggregate Cultures (MAAC)

DOI: 10.3791/60485-v
What you'll learn
  • ✓ Isolate mature human adipocytes from subcutaneous adipose tissue
  • ✓ Culture mature adipocytes using the MAAC method
  • ✓ Assess adipocyte viability and pharmacological responses in culture
Protocol

Membrane mature adipocyte aggregate culture (MAAC) is a new method to culture mature human adipocytes. Here we detail how to isolate adipocytes from human adipose and how to set up MAAC.

Difficulty
advanced
Total time
~4–6 hours per sample (tissue dissection through culture setup)
Model organism
Human adipose tissue (primary mature adipocytes)
Biosafety
BSL-1

Steps

1
Introduce MAAC method and experimental overview

Review the membrane mature adipocyte aggregate culture (MAAC) technique and its advantages for culturing human mature adipocytes ex vivo.

▶ 00:05
2
Dissect human subcutaneous adipose tissue

Obtain and prepare human subcutaneous adipose tissue sample for enzymatic digestion by careful dissection and removal of connective tissue.

▶ 00:53
3
Treat adipose tissue with collagenase enzyme

Incubate dissected adipose tissue with collagenase to digest the extracellular matrix and release mature adipocytes from the tissue.

▶ 01:32
4
Purify mature adipocytes from stromal cells

Separate mature adipocytes from stromal vascular fraction through flotation or density-based separation to obtain a pure adipocyte population.

▶ 02:21
5
Pack adipocytes for aggregate culture formation

Concentrate and pack purified mature adipocytes into a defined configuration to establish the three-dimensional aggregate structure for MAAC.

▶ 03:20
6
Seed and maintain mature adipocyte aggregates

Inoculate MAAC constructs into culture medium and maintain under appropriate conditions to support long-term adipocyte viability and metabolic function.

▶ 03:51
7
Evaluate MAAC appearance and pharmacological responses

Assess morphology of mature adipocyte aggregate cultures and measure cellular responses to pharmacological agonists to validate culture functionality.

▶ 04:50
8
Summarize MAAC protocol and applications

Review key findings and discuss the utility of MAAC for studying mature human adipocyte biology and drug responses.

▶ 05:55
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