Method for cultivating stem cells in vitro
AssigneeSimone Spuler
InventorsAndreas Marg, Simone Spuler
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Abstract
The invention relates to method for cultivating stem cells in vitro, comprising the following steps: providing a sample comprising stem cells and cultivating the stem cells by subjecting the sample to a treatment for a first period of time. The treatment is carried out under hypothermic conditions having a defined temperature and a defined atmosphere, wherein the temperature does not exceed 15° C. and the atmosphere has an oxygen content not exceeding 21% (v/v). Thereby, the first period of time is 4 days to 4 weeks.
Claims12
A method for generating genetically modified human muscle satellite cells, the method comprising the steps of:
- obtaining a human muscle biopsy specimen from a living subject;
- mechanically isolating, without enzymatic treatment, muscle fiber fragments from the biopsy specimen to provide isolated muscle fiber fragments comprising fiber fragments of a length of 2 mm to 3 mm;
- incubating the isolated muscle fiber fragments at about 4° C. to about 8° C. for a period of 5 days to 7 days;
- immediately after incubation, culturing the isolated muscle fiber fragments at about 37° C. under conditions sufficient for satellite cells present in the isolated muscle fiber fragments to form cell colonies that outgrow from the muscle fiber fragments;
- and
- collecting the satellite cells from the cell colonies, wherein 70% to 100% of viable cells collected from the cell colonies are satellite cells, and
- inserting genetic information into the genome of the satellite cells or replacing defect genetic information with desired genetic information.
Dependencies are inferred from the claim text.