US 9,790,469 B2 · Established human brown adipocyte line and method for differentiation from an hMADS cell line

Abstract

The subject matter of the invention is a functional population of human brown adipocytes, in which the expression of UCP1, CIDEA, CPT1B and Bc12 is higher, the expression of Bax is lower and the expression of PPAR-alpha, PGC-1alpha, PGC-1 beta and PRDM16 is similar compared with the corresponding expressions of a population of human white adipocytes. The invention also relates to a method for differentiation of hMADS cells into the functional population of human brown adipocytes, to a method for conversion of a population of human white adipocytes into the functional population of human brown adipocytes, and also to a method of screening for molecules capable of modulating the bodyweight in an individual.

Claims18

Dependent claims

A method for differentiating a population of human multipotent adipose-derived stem cells (hMADS cells) into a functional human brown adipocytes population, comprising the steps of:

a) culturing the hMADs cells in a first differentiation medium consisting essentially of nutrients, transferrin, dexamethasone (DEX), isobutylmethylxanthine (IBMX), insulin, and T3 for a duration period comprised between 2 and 4 days, thereby initiating the differentiation of the hMADs cells;
and
b) culturing the cells from step a) in a second differentiation medium comprising nutrients, transferrin, insulin, T3 and a specific PPAR gamma agonist for 30 days, thereby differentiating the cells from step a) into functional human brown adipocytes,
wherein the second differentiation medium does not comprise DEX or IBMX.

1 more element

A method for converting a human white adipocytes population into a functional human brown adipocytes population, comprising the steps of:

a) culturing said human white adipocytes population in a differentiation medium comprising nutrients, transferrin, insulin, T3 and a specific PPAR gamma agonist, for a duration period comprised between 1 and 10 days, thereby converting the human white adipocytes into functional human brown adipocytes,
wherein the differentiation medium does not comprise dexamethasone (DEX) or isobutylmethylxanthine (IBMX).

A method for differentiating a population of human multipotent adipose-derived stem cells (hMADS cells) into a functional human brown adipocytes population, comprising the steps of:

a) culturing the hMADs cells in a first differentiation medium comprising dexamethasone (DEX), isobutylmethylxanthine (IBMX), insulin, and T3 for a duration period comprised between 2 and 4 days, thereby initiating the differentiation of the hMADs cells;
b) culturing the cells from step a) in a second differentiation medium comprising insulin, T3 and a first specific PPAR gamma agonist for a duration period comprised between 2 and 9 days, thereby differentiating the cells from step a) into white adipocytes;
c) culturing the white adipocytes from step b) for a duration period comprised between 2 to 10 days without the stimulation of differentiation with a PPAR gamma agonist, and
d) culturing the cells from step c) for a duration period comprised between 1 and 10 days, in a third differentiation medium comprising insulin, T3 and a second specific PPAR gamma agonist, thereby differentiating the cells from step c) into functional human brown adipocytes,
wherein the first and second PPARγ gamma agonists may be the same or different, and
wherein the second differentiation medium and the third differentiation medium do not comprise the DEX or the IBMX.

3 more elements

Dependencies are inferred from the claim text.

Established human brown adipocyte line and method for differentiation from an hMADS cell line

Life of this patent

Established human brown adipocyte line and method for differentiation from an hMADS cell line

Life of this patent

Abstract

Claims18