Decellularized Adipose Cell Growth Scaffold

Abstract

Provided herein are methods of making a cell growth scaffold from adipose tissue, cell growth scaffolds having low lipid content and methods of using the cell growth scaffold.

We claim: 
     
         1 . A cell growth scaffold prepared by washing a decellularized adipose tissue-derived cell growth scaffold with n-propanol, isopropanol or a mixture thereof. 
     
     
         2 . The cell growth scaffold of  claim 1 , prepared by, in order: digesting an adipose tissue sample with a proteinase and a chelating agent; washing the adipose tissue sample with an emulsifier; disinfecting the adipose tissue sample; depyrogenating the adipose tissue sample; drying the adipose tissue sample; and washing the adipose tissue sample with n-propanol, isopropanol or a mixture thereof. 
     
     
         3 . The cell growth scaffold of  claim 2 , prepared by a method comprising the following steps, in order:
 a. rinsing an adipose tissue sample;   b. mechanically massaging the adipose tissue sample;   c. digesting the adipose tissue sample with a proteinase and a chelating agent;   d. rinsing the adipose tissue sample;   e. mechanically massaging the tissue sample;   f. washing the adipose tissue sample with a surfactant while the adipose tissue sample is agitated;   g. rinsing the adipose tissue sample;   h. washing the adipose tissue sample with an emulsifier;   i. rinsing the adipose tissue sample;   j. disinfecting the adipose tissue sample;   k. depyrogenating the adipose tissue sample;   l. rinsing the adipose tissue sample;   m. drying the adipose tissue sample;   n. washing the dried adipose tissue sample in n-propanol, isopropanol or a mixture thereof;   o. washing the adipose tissue sample in an aqueous solvent; and   p. drying the adipose tissue sample.   
     
     
         4 . A method of growing cells comprising contacting cells with a cell growth scaffold prepared by washing a decellularized adipose tissue-derived cell growth scaffold with n-propanol, isopropanol or a mixture thereof. 
     
     
         5 . The method of  claim 4 , wherein the cell growth scaffold is prepared by, in order: digesting an adipose tissue sample with a proteinase and a chelating agent; washing the adipose tissue sample with an emulsifier; disinfecting the adipose tissue sample; depyrogenating the adipose tissue sample; drying the adipose tissue sample; and washing the adipose tissue sample with n-propanol, isopropanol or a mixture thereof. 
     
     
         6 . The method of  claim 5 , wherein the cell growth scaffold is prepared by a method comprising the following steps, in order:
 a. rinsing an adipose tissue sample;   b. mechanically massaging the adipose tissue sample;   c. digesting the adipose tissue sample with a proteinase and a chelating agent;   d. rinsing the adipose tissue sample;   e. mechanically massaging the tissue sample;   f. washing the adipose tissue sample with a surfactant while the adipose tissue sample is agitated;   g. rinsing the adipose tissue sample;   h. washing the adipose tissue sample with an emulsifier;   i. rinsing the adipose tissue sample;   j. disinfecting the adipose tissue sample;   k. depyrogenating the adipose tissue sample;   l. rinsing the adipose tissue sample;   m. drying the adipose tissue sample;   n. washing the dried adipose tissue sample in n-propanol, isopropanol or a mixture thereof;   o. washing the adipose tissue sample in an aqueous solvent; and   p. drying the adipose tissue sample.   
     
     
         7 . The method of  claim 4 , wherein the cells are cultured in vivo. 
     
     
         8 . The method of  claim 4 , wherein the cell growth scaffold is applied to a wound in a patient. 
     
     
         9 . The method of  claim 4 , wherein the cell growth scaffold is applied to a topical wound. 
     
     
         10 . The method of  claim 9 , wherein the cell growth scaffold is sprayed onto a topical wound. 
     
     
         11 . The method of  claim 4 , wherein the scaffold is contained within a biocompatible cover to produce an implant, and the implant is implanted into a patient. 
     
     
         12 . The method of  claim 11 , wherein the implant comprises cells. 
     
     
         13 . The method of  claim 12 , wherein the cells are chondrocytes, chondroprogenitor cells, adipose stem cells, mesenchymal stem cells, myocytes, precursor cells, progenitor cells, stem cells, differentiated cells. 
     
     
         14 . The method of  claim 4 , wherein the scaffold is surgically implanted into a patient. 
     
     
         15 . The method of  claim 4 , wherein the scaffold is injected into a patient. 
     
     
         16 . A method of treating a wound or defect in a patient, comprising injecting or surgically implanting into a patient a cell growth scaffold prepared by washing a decellularized adipose tissue-derived cell growth scaffold with n-propanol, isopropanol or a mixture thereof. 
     
     
         17 . The method of  claim 16 , wherein the cell growth scaffold is prepared by, in order: digesting an adipose tissue sample with a proteinase and a chelating agent; washing the adipose tissue sample with an emulsifier; disinfecting the adipose tissue sample; depyrogenating the adipose tissue sample; drying the adipose tissue sample; and washing the adipose tissue sample with n-propanol, isopropanol or a mixture thereof. 
     
     
         18 . The method of  claim 17 , wherein the cell growth scaffold is prepared by a method comprising the following steps, in order:
 a. rinsing an adipose tissue sample;   b. mechanically massaging the adipose tissue sample;   c. digesting the adipose tissue sample with a proteinase and a chelating agent;   d. rinsing the adipose tissue sample;   e. mechanically massaging the tissue sample;   f. washing the adipose tissue sample with a surfactant while the adipose tissue sample is agitated;   g. rinsing the adipose tissue sample;   h. washing the adipose tissue sample with an emulsifier;   i. rinsing the adipose tissue sample;   j. disinfecting the adipose tissue sample;   k. depyrogenating the adipose tissue sample;   l. rinsing the adipose tissue sample;   m. drying the adipose tissue sample;   n. washing the dried adipose tissue sample in n-propanol, isopropanol or a mixture thereof;   o. washing the adipose tissue sample in an aqueous solvent; and   p. drying the adipose tissue sample.   
     
     
         19 . The method of  claim 16 , wherein the scaffold is contained within a biocompatible cover to produce an implant. 
     
     
         20 . The method of  claim 16 , wherein the implant comprises cells.