Coatings and culture media for promoting neurogenesis in adipose tissue derived stem cells

Patent metadata

Field	Value
Publication number	US-9428730-B2
Application number	US-201213408490-A
Country	US
Kind code	B2
Filing date	Feb 29, 2012
Priority date	Feb 29, 2012
Publication date	Aug 30, 2016
Grant date	Aug 30, 2016

Abstract

Provided herein are coatings for stem cell cultureware comprising poly-L-ornithine and bovine fibronectin and methods for preparing coated stem cultureware comprising contacting cultureware with poly-L-ornithine and contacting the cultureware with bovine fibronectin. Also provided are stem cell culture media comprising epidermal growth factor, beta-fibroblast growth factor and N 2 supplement.

First claim

We claim:

1. A neuronal differentiation medium for culturing human adipose derived stem cells consisting of:
a neural basal medium, wherein the neural basal medium consists of glycine, L-alanine, L-arginine, L-asparagine, L-cysteine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, choline, D-calcium pantothenate, folic acid, niacinamide, pyridoxine hydrochloride, riboflavin, thiamine hydrochloride, vitamin B12, i-inositol, calcium chloride, ferric nitrate, magnesium chloride, potassium chloride, sodium bicarbonate, sodium chloride, sodium phosphate, zinc sulfate, dextrose, 4-(Z-hydroxyethyl)-1-piperazineethanesulfonic acid, sodium pyruvate,
epidermal growth factor (EGF) at a concentration about 1 to about 100 ng/mL, b-fibroblast growth factor (b-FGF) at a concentration of about 1 to 100 ng/mL, and N2 supplement at a concentration of about 1x, wherein the medium promotes neurogenesis, and wherein the neuronal differentiation medium is substantially free of serum.

2. The medium of claim 1 , wherein the N2 supplement comprises human transferrin, full chain recombinant insulin, progesterone, putrescine, and selenite.

3. The medium of claim 1 , wherein the medium promotes neurogenesis by inducing one or more morphological changes in one or more cells exposed to the medium, wherein the one or more morphological changes observed include at least one selected from the group consisting of shrinkage of the cytoplasm, promoting neurite formation, promoting axon formation, promoting dendrite formation, and combinations thereof.

4. The medium of claim 3 , wherein the one or more cells exposed to the medium are adipocyte derived stem cells.

5. The medium of claim 4 , wherein the one or more cells exposed to the medium are human adipocyte derived stem cells.

6. A neuronal differentiation medium for culturing human adipose derived stem cells consisting of:
a neural basal medium, wherein the neural basal medium consists of from the group consisting of glycine, L-alanine, L-arginine, L- asparagine, L-cysteine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, choline, D-calcium pantothenate, folic acid, niacinamide, pyridoxine hydrochloride, riboflavin, thiamine hydrochloride, vitamin B12, i-inositol, calcium chloride, ferric nitrate, magnesium chloride, potassium chloride, sodium bicarbonate, sodium chloride, sodium phosphate, zinc sulfate, dextrose, 4-(Z-hydroxyethyl)-1-piperazineethanesulfonic acid, sodium pyruvate,
epidermal growth factor (EGF) at a concentration about 1 to about 100 ng/mL, b-fibroblast growth factor (b-FGF) at a concentration of about 1 to 100 ng/mL, N2 supplement at a concentration of about 1x, and L-glutamine at a concentration of about 0.1 to about 10 mM, wherein the medium promotes neurogenesis, and wherein the neuronal differentiation medium is substantially free of serum.

7. The medium of claim 6 , wherein the N2 supplement comprises human transferrin, full chain recombinant insulin, progesterone, putrescine, and selenite.

8. The medium of claim 6 , wherein the medium promotes neurogenesis by inducing one or more morphological changes in one or more cells exposed to the medium, wherein the one or more morphological changes observed include at least one selected from the group consisting of shrinkage of the cytoplasm, promoting neurite formation, promoting axon formation, and promoting dendrite formation.

9. The medium of claim 8 , wherein the one or more cells exposed to the medium are adipocyte derived stem cells.

10. The medium of claim 9 , wherein the one or more cells exposed to the medium are human adipocyte derived stem cells.

Assignees

• Kuang Chenzhong
• Xiao Yan
• Jouni Zeina
• Poels Eduard K
• Hondmann Dirk
• Mead Johnson Nutrition Co

Inventors

• Kuang Chenzhong
• Xiao Yan
• Jouni Zeina
• Poels Eduard K
• Hondmann Dirk

Classifications

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N2533/52

  Fibronectin; Laminin · CPC title

• C12N5/0618 Primary

  Cells of the nervous system · CPC title

• C12N2533/32

  Polylysine, polyornithine · CPC title

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title