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. Author manuscript; available in PMC: 2016 Nov 1.
Published in final edited form as: Biotechnol Adv. 2015 Jul 29;33(6 0 1):962–979. doi: 10.1016/j.biotechadv.2015.07.005

Table 2.

Stem Cell Differentiation into Brown Adipocytes in vitro

Stem Cell Population Differentiation Scheme Results References
Mouse MSCs

  1. C3H10T1/2 cell growth with BMP7 until confluence (3 days)

  2. Brown adipocyte differentiation: cells exposed to adipogenic cocktail for 48 hours and then placed into growth medium containing insulin and T3 (4–5 more days)

  • Exhibited multilocular lipid formation and UCP1 expression;

  • Mitochondrial biogenesis indicated by PGC1α, NRF1, TFAM, and cytochrome C expression at expected timepoints.

 Tseng et al., 2008
hASCs

  1. hMADS-2 cell growth with media containing hFGF2 until confluence

  2. Brown adipocyte differentiation: 2 days post-confluence cells treated with an adipogenic cocktail (3 days) and then treated with a medium containing rosiglitazone with dexamethasone and IBMX removed (up to 17 more days)

  • Increased UCP1, CIDEA, and CPT1B expression and increased total respiration and oligomycin-sensitive respiration

  • Increased UCP1 expression in response to isoproterenol and CL316243 (β- and β3-agonists, respectively)

  • Responsive to β1 and β3 agonists but not β2

 Elabd et al., 2009
Mattsson et al., 2011
Pisani et al., 2011
hESCs

  1. EB formation: hESCs + growth medium in suspension with growth medium (7 days)

  2. Mesenchymal progenitor cell (MPC) formation: replated EB’s + growth medium until confluency (5 days) and then replated + MPC growth medium. MPCs were split 1:3 prior to further differentiation.

  3. Brown adipocyte formation: lentiviral transduction of MPCs containing combinations of PPARγ2, C/EBPB, and PRDM16 + adipogenic differentiation medium with (14 days) and without doxycycline (until day 21 or later)

  • More greatly induced UCP1 by PPARγ2-C/EBPβ and PPARγ2-C/EBPβ-PRDM16 than C/EBPβ-PRDM16

  • Multilocular lipid accumulation in programmed brown adipocytes in both PPARγ2-containing transcription factor combinations

  • Exhibited functional properties with high oxygen consumption and extracellular acidification rates

 Ahfeldt et al., 2012

Lee and Cowan, 2014
hiPSCs

  1. Sphere formation: hiPSCs + differentiation medium + hematopoietic cytokine cocktail I (8 days)

  2. Brown adipocyte formation: replated spheres + differentiation medium + hematopoietic cytokine cocktail II (including BMP7 for several days)

  • Induced brown adipocyte differentiation by hematopoietic cocktail regardless of BMP7

  • Formed multilocular lipid droplets

  • Increased UCP1 and PRDM16 expression upon β-adrenergic receptor agonist

 Nishio et al., 2012
hiPSCs

  1. EB formation: hiPSCs in suspension (10 days, with and without RA added from days 3–5)

  2. MSC formation: replated EB outgrowths + mesenchymal cell growth medium (7 days)

  3. Adipocyte progenitor (APs) formation: collection and expansion of cells being able to form clones 10 days after replating

  4. Brown AP formation: APs transduced with C/EBPβ/LAP-expressing retroviral vector + induced with adipogenic medium

  • Increased UCP1, Dio2, and PGC1α expression upon β-adrenergic stimulation

  • Resulted in low UCP1 expression by RA treatment prior to brown AP formation

  • Displayed higher clonogenic potential by brown APs than white APs

  • Enriched Pax3 expression in brown APs compared with white APs

 Cedex et al., 2014