Apparatus for Enhanced Recovery of Regenerative cells From Tissue Samples

Abstract

This document describes methods and an apparatus for recovery of a cell-enriched matrix and cells (e.g., regenerative cells) from a tissue sample. In some embodiments, at least two rounds of acceleration and deceleration are performed.

1 . A cellular composition comprising the combination of (i) a cell-enriched matrix prepared from a sample of body tissue having vasculature and regenerative cells adaptive to differentiate into functional equivalent of host tissue after transplantation therein, by extruding the body tissue sample through an orifice to reduce lipid content and increase regenerative cell content in the sample, followed by centrifugation of the extruded sample at a centrifugal force and duration sufficient to derive therefrom the cell-enriched matrix with regenerative platform therein; together with (ii) a regenerative cell population recovered by subjecting such a derived cell-enriched matrix to repetitive cycles of rotational acceleration and deceleration of at least one cycle thereof per minute in a temperature-controlled centrifuge with enzymatic environment to dissociate and extract regenerative cells from the matrix. 
     
     
         2 . The composition of  claim 1  wherein the body tissue is selected from one of lipoaspirate, adipose tissue, and combinations thereof. 
     
     
         3 . The composition of  claim 1  wherein centrifugation of the extruded sample is applied with a force in a range from about 400 g to about 2000 g for a duration of about 5 minutes. 
     
     
         4 . The composition of  claim 1  wherein the orifice through which the sample of body tissue is extruded has a diameter in a range from about 1 mm to about 5 mm. 
     
     
         5 . The composition of  claim 1  wherein the rotational acceleration and deceleration is performed by securing such a cell-derived matrix in a container to an inverted rotor of a centrifuge, and subjecting the secured container to a sequence of multiple rounds of the rotational acceleration and deceleration of at least one round per minute at a centrifugal force in a predetermined range for a duration sufficient for the dissociation and extraction of regenerative cells. 
     
     
         6 . The composition of  claim 5  wherein the multiple cell-enriched matrix containers are secured radially in spaced-apart orientation to the rotor for retention thereof at one of (i) a fixed angle or (ii) an angle that varies with rotational force on the respective container, and the rotor is spun under rotational forces through the sequential rounds of rotational acceleration and deceleration at a rate of said at least one round per minute, to dissociate regenerative cells from the contained cell-enriched matrix as a combined result of conflicting centrifugal and gravitational forces thereon. 
     
     
         7 . The composition of  claim 6  wherein said angle, whether fixed or varying with rotational forces on the container, is positively acute between and including a horizontal plane through the rotational axis of the rotor and a vertical orientation of said axis so as to create said conflicting centrifugal and gravitational forces during start-stop cycles resulting from said sequential rounds of rotational acceleration and deceleration. 
     
     
         8 . The composition of  claim 1  wherein the enzymatic environment in said temperature controlled centrifuge is provided by a proteolytic enzyme selected from a collagenase, a neutral protease, or a combination thereof. 
     
     
         9 . The composition of  claim 5  wherein the temperature within said temperature controlled centrifuge is maintained in a range from about 26° C. to about 42° C. during said multiple rounds of rotational acceleration and deceleration. 
     
     
         10 . The composition of  claim 5  wherein the sequential rounds of rotational acceleration and deceleration are substantially uninterrupted during centrifugation. 
     
     
         11 . The composition of  claim 5  wherein the cellular composition is suitable to be banked as is or in a resorbable scaffold, for subsequent implantation into a subject. 
     
     
         12 . The composition of  claim 11  wherein the banking is enabled by first subjecting the cellular composition or scaffold to cryopreservation. 
     
     
         13 . The composition of  claim 1  wherein said cell-enriched matrix and said regenerative cell population exhibits enhanced neovascularization for long term survival as a transplanted volume-filling graft. 
     
     
         14 . The composition of  claim 1  prepared for implantation in the same subject from which said sample was originally taken. 
     
     
         15 . The composition of  claim 11  wherein the subject to receive the implantation is different from the subject from which said sample was originally acquired. 
     
     
         16 . The composition of  claim 10  wherein the cellular composition possesses a capability to restore cellular function of non-functioning tissue of a subject when implanted at a target site of the non-functioning tissue.