Culture media for multipotent stem cells
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Publication Number
US-11111480-B2
Publication Date
2021-09-07
Expiration Date
Abstract
A growth medium for culturing mesenchymal stem cells. The medium comprises a serum, a fibroblast growth factor, and either an L-cysteine, a glutathione, or an N-acetyl cysteine. Optionally, the medium can comprise various quantities of an epidermal growth factor, a hydrocortisone, a calcium chloride, an insulin, a pituitary extract, a selenium, a stromal-derived factor, a sodium pyruvate, a transferrin, and serum-free medium.
Core Innovation
The disclosed invention relates to a mesenchymal stem cell growth medium comprising serum, a fibroblast growth factor, and one of L-cysteine, glutathione, or N-acetyl cysteine. The medium further includes epidermal growth factor, hydrocortisone, calcium chloride, insulin, pituitary extract, selenium, stromal-derived factor, sodium pyruvate, transferrin, and serum-free medium balanced to 100 percent volume.
The invention states that the mesenchymal stem cell growth medium preserves mesenchymal stem cell characteristics over time. The disclosure includes example component proportions and evaluates outcomes using morphology, proliferation, multilineage differentiation, and immunophenotype results.
Claims Coverage
The independent claim defines a mesenchymal stem cell growth medium with multiple specified components and concentration ranges, together with a functional requirement that cultured mesenchymal stem cells are morphologically identical to mesenchymal stem cells of a previous generation.
Morphologically identical cultured mesenchymal stem cells across generations
A requirement wherein a cultured mesenchymal stem cell is morphologically identical to a mesenchymal stem cell of a previous generation.
Specified multi-component mesenchymal stem cell growth medium ranges
A growth medium for culturing mesenchymal stem cells comprising a serum from about 1 percent to about 10 percent by volume, a fibroblast growth factor from about 1 ng/mL to about 10 ng/mL, either L-cysteine from about 0.5 mM to about 5 mM or glutathione from about 0.5 mM to about 5 mM, an epidermal growth factor from about 1 ng/ml to about 10 ng/ml, hydrocortisone from about 10 ng/mL to about 100 ng/mL, calcium chloride from about 0.01 mM to about 0.1 mM, insulin from about 0.5 mg/mL to about 5 mg/mL, pituitary extract from about 10 μg/mL to about 100 μg/mL, selenium from about 0.1 μg/mL to about 1 μg/mL, stromal-derived factor from about 1 ng/mL to about 10 ng/mL, sodium pyruvate from about 2 mg/mL to about 20 mg/mL, transferrin from about 0.1 mg/mL to about 1 mg/mL, and serum free medium balanced to 100 percent volume.
Human serum as the serum component
The stem cell growth medium uses human serum as the serum component.
Fetal bovine serum as an alternative serum source
The stem cell growth medium uses fetal bovine serum as the serum component.
Recombinant fibroblast growth factor and/or recombinant epidermal growth factor
At least one of the fibroblast growth factor or epidermal growth factor is recombinant.
Recombinant stromal-derived factor
The stem cell growth medium is characterized by using recombinant stromal-derived factor as the stromal-derived factor.
L-cysteine as the sulfur-containing component alternative
The stem cell growth medium contains L-cysteine.
Overall claim coverage is centered on a defined mesenchymal stem cell growth medium with specified concentration ranges and multiple additives, combined with a morphological preservation requirement across generations. Dependent claims further restrict particular selections, including serum source (human or fetal bovine), recombinant forms of selected growth factors, a recombinant stromal-derived factor, and selection of L-cysteine as the specified sulfur-containing alternative.
Stated Advantages
Preserves mesenchymal stem cell characteristics over time.
Documented Applications
Not explicitly described in patent.
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