Method of making three-dimensional, leaf-based scaffold for three-dimensional cell culture

Patent metadata

Field	Value
Publication number	US-9783777-B1
Application number	US-201615297096-A
Country	US
Kind code	B1
Filing date	Oct 18, 2016
Priority date	Oct 18, 2016
Publication date	Oct 10, 2017
Grant date	Oct 10, 2017

Abstract

The method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures includes washing a quantity of Ficus religiosa leaves, then treating the washed Ficus religiosa leaves in a sodium hydroxide solution to obtain alkali-treated Ficus religiosa leaves. The alkali-treated Ficus religiosa leaves are washed, and then superficial tissue is removed from the alkali-treated Ficus religiosa leaves to obtain Ficus religiosa leaf skeletons. The Ficus religiosa leaf skeletons are dried and then consecutively immersed in distilled water, a phosphate buffer saline solution, and plain Dulbecco's modified Eagle's medium (DMEM) to form the three-dimensional scaffolds for three-dimensional cell cultures. Each three-dimensional scaffold can be used for growing three-dimensional cell cultures, such as human mesenchymal stem cell cultures.

First claim

We claim:

1. A method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures, comprising the steps of:
treating Ficus religiosa leaves in a sodium hydroxide solution to obtain alkali-treated Ficus religiosa leaves;
washing the alkali-treated Ficus religiosa leaves;
removing layers of superficial tissue from the alkali-treated Ficus religiosa leaves to obtain Ficus religiosa leaf skeletons;
drying the Ficus religiosa leaf skeletons to obtain dried Ficus religiosa leaf skeletons; and
immersing the dried Ficus religiosa leaf skeletons in a cell culture medium to provide a three-dimensional, leaf-based scaffold for three-dimensional cell cultures, wherein the cell culture medium is Dulbecco's modified Eagle's medium (DMEM).

2. The method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures as recited in claim 1 , wherein the sodium hydroxide solution comprises in a 4.0 vol % aqueous solution of sodium hydroxide.

3. The method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures as recited in claim 2 , further comprising heating the washed Ficus religiosa leaves at a temperature of approximately 120° C. for approximately 60 minutes under a pressure of approximately 15 lbs.

4. The method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures as recited in claim 1 , wherein the step of drying the Ficus religiosa leaf skeletons comprises drying the Ficus religiosa leaf skeletons at a temperature of approximately 50° C. for approximately 30 minutes.

5. The method of making a three-dimensional, leaf-based scaffold for three-dimensional cell cultures as recited in claim 1 , further comprising:
consecutively immersing the dried Ficus religiosa leaf skeletons in distilled water and a phosphate buffer saline solution, prior to the step of immersing the dried Ficus religiosa leaf skeletons in the cell culture medium.

Assignees

• Univ King Saud

Inventors

• Subbarayan Periasamy Vaiyapuri
• Alshatwi Ali Abdullah
• Athinarayanan Jegan

Classifications

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• C12N5/0062 Primary

  General methods for three-dimensional culture · CPC title

• C12N2513/00

  3D culture · CPC title

• C12N5/0668

  Mesenchymal stem cells from other natural sources · CPC title

• C12N2500/76

  from plants · CPC title