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Brown fat cell compositions and methods

A technology of brown adipose tissue and brown fat, which is applied in the field of stem cell culture and application, and can solve the problem of identifying stem cell populations without brown adipose tissue

Inactive Publication Date: 2014-07-16
BIORESTORATIVE THERAPIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To date, no population of stem cell populations have been identified in brown adipose tissue

Method used

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  • Brown fat cell compositions and methods
  • Brown fat cell compositions and methods
  • Brown fat cell compositions and methods

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Embodiment 1-brown fat separation processing

[0118] In one embodiment, the patient first experiences 18 F-fluorodeoxyglucose ( 18 F-FDG) PET-CT scan to identify brown fat deposits. Before undergoing the PET-CT scan, the patient was exposed to a temperature range of 1°C-25°C daily (to increase the 18 Ingestion of F-FDG) for 1-2 hours for a month. Using a needle, biopsies were taken of identified deposits in the supraclavicular and mediastinal regions. Brown fat biopsies were washed three times in DPBS(-) and processed with 0.075%-0.2% collagenase type 1A for 30-60 minutes at 37°C with vigorous shaking or with EDTA for 30 minutes and subjected to 5-30 minutes of incubation. Mechanical tissue destruction. Tissues / cells were washed three times with DPBS(-), and 1x10 6 ~1x10 10 cells were injected into the patient's white fat deposits. Optionally, after enzymatic or mechanical tissue / cell dissociation, the tissue / cells are exposed to cold temperature (0°C-4°C) fo...

Embodiment 2-

[0119] Example 2 - Brown Fat Culture

[0120] In this example, the cells are cultured prior to injection into the patient. The cells obtained from Example 1 were used at 1000 cells / cm 2 The concentration of is inoculated (plate) into the culture medium of the super flask (hyperflask). In this embodiment, the culture medium contains:

[0121] DMEM, low glucose, without phenol red;

[0122] 0.5-20% human platelet lysate;

[0123] 1X NEAA;

[0124] 1X Glutamax;

[0125] 1X Gentamicin;

[0126] 1000 units of heparin.

[0127] The cells were cultured for 3-6 weeks. The cells were then characterized using flow cytometry and analyzed for expression of one or more of the following markers: SCA-1, CD34, SSEA1, SSEA4, OCT4, CD31, Wnt5a, telomerase activity, α-SMA, STRO-1, MYF5, PRDM16 or UCP1. The cells were then expanded to 1x10 6 ~1x10 10 concentration to be implanted into the patient's white fat deposits.

Embodiment 3

[0128] Example 3 - Differentiation of cells to brown adipocytes

[0129] In this embodiment, cells obtained from brown adipose deposits of a patient (such as stem cells or precursor brown adipocytes) are differentiated and then injected into the patient. The cells obtained from Example 1 were used at 1000 cells / cm 2 Inoculate into a superflask containing differentiation medium, which in this embodiment comprises:

[0130] DMEM, low glucose, without phenol red;

[0131] 0.5-20% human platelet lysate;

[0132] 1X NEAA;

[0133] 1X Glutamax;

[0134] 1X Gentamicin;

[0135] 1000 units of heparin;

[0136] 10 μg / mL insulin;

[0137] 1 μM dexamethasone;

[0138] 200μM indomethacin;

[0139] 0.5mM isobutylmethylxanthine;

[0140] In an alternative embodiment, the differentiation medium comprises:

[0141] DMEM (low glucose, without phenol red);

[0142] 0.5%-20% human platelet lysate;

[0143] 1X NEAA; 1X Glutamax;

[0144] 1X Gentamicin;

[0145] 1000 units of hepa...

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Abstract

Methods of developing and using cell lines, such as stem cell lines, for therapeutic or cosmetic use. In one embodiment the cell lines are used to treat a wide range of degenerative and metabolic disorders including, but not limited to, obesity, diabetes, hypertension, and cardiac deficiency. Also described are methods of using such cell lines to screen for compounds that play a role in regulating a variety of processes.

Description

technical field [0001] The present invention relates to the field of cell culture and more particularly to the culture and use of stem cells. [0002] related application [0003] This application claims priority to U.S. Provisional Application 61 / 502,508, filed June 29, 2011, U.S. Provisional Application 61 / 632,122, filed January 18, 2012, and U.S. Provisional Application 61 / 632,516, filed January 25, 2012 and benefits, each of which is hereby incorporated by reference in its entirety. Background technique [0004] Brown fat (brown fat) is one of two types of adipose tissue found in the human body. Brown fat is derived from the differentiation of mesoderm during embryogenesis. Brown fat is involved in development and homeostasis by providing metabolic tissue capable of providing heat. Brown fat helps regulate metabolism and nonshivering thermogenesis. In addition, brown fat plays a greater role in regulating metabolism than white adipose tissue. Brown fat makes up 5 p...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775A61K35/28
CPCA61K35/28C12N2500/84C12N2500/98C12N2501/01C12N2501/02C12N2501/33C12N2501/385C12N2501/39C12N2501/395C12N2506/1384C12N5/0653C12N5/0667A61P3/04Y02A50/30A61K35/35G01N33/502G01N33/5044
Inventor 弗朗西斯科·哈维尔·席尔瓦马克·魏因雷布阿米特·N·帕特尔戴维·A·布尔
Owner BIORESTORATIVE THERAPIES
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