Compositions for biological systems and methods for preparing and using the same

Abstract

A method of making a composition for influencing biological growth includes obtaining a sample comprising human umbilical cord blood and having a first volume VUC, combining the sample with a quantity of PrepaCyte-CB or the equivalent having a volume VPC, wherein the ratio VUC/VPC is between about 0.60 and about 1.75 such that a supernatant is formed, layering within a container at least a portion of the supernatant over a layer of a reagent configured for isolating mononuclear cells, the at least a portion of the supernatant having a volume VS and the layer of the reagent having a volume VR, wherein the volume of the layer of the reagent VR is at least 67% the volume VS of the at least a portion of the supernatant, placing a sufficient centrifugal force on the contents of the container to cause at least a new layer substantially comprising mononuclear cells to form, combining at least some of the layer substantially comprising mononuclear cells with lactated Ringer's solution or the equivalent to create a mononuclear cell solution, determining the total number of live cells NL in at least a portion of the mononuclear cell solution, the at least a portion of the mononuclear solution having a volume VM, and adding a volume VD of Low Molecular Weight Dextran in Dextrose solution to the at least a portion of the mononuclear cell solution, the volume VD based at least in part on the total number of live cells NL.

What is claimed is: 
     
         1 . A composition for performing a medical therapy, comprising:
 a low pH fluid base having a volume V DD ;   live cells obtained from human umbilical cord blood; and   a protectant,   wherein the volume V DD  is a result of predicted viability following freezing and thawing of the live cells as combined with the low pH fluid base and the protectant, and wherein the composition is configured for implantation within a human subject.   
     
     
         2 . The composition of  claim 1 , wherein the low pH fluid base comprises dextran. 
     
     
         3 . The composition of  claim 2 , wherein the low pH fluid base comprises dextrose. 
     
     
         4 . The composition of  claim 1 , wherein the low pH fluid base comprises dextrose. 
     
     
         5 . The composition of  claim 1 , wherein the live cells comprise mononuclear cells. 
     
     
         6 . The composition of  claim 5 , wherein the mononuclear cells are present in the composition at between about 100,000 cells per ml and about 75 million cells per ml. 
     
     
         7 . The composition of  claim 5 , wherein the mononuclear cells are derived only from umbilical cord blood of a single donor. 
     
     
         8 . The composition of  claim 1 , wherein the protectant comprises albumin. 
     
     
         9 . The composition of  claim 1 , wherein the protectant comprises a cryoprotectant. 
     
     
         10 . The composition of  claim 1 , wherein the protectant does not comprise dimethyl sulfoxide (DMSO). 
     
     
         11 . The composition of  claim 1 , wherein the predicted viability within previously living cells was approximately 75%. 
     
     
         12 . The composition of  claim 1 , further comprising one or more bioactive agents. 
     
     
         13 . The composition of  claim 12 , wherein the one or more bioactive agents comprise one or more cytokines. 
     
     
         14 . The composition of  claim 12 , wherein the one or more bioactive agents comprise one or more growth factors. 
     
     
         15 . The composition of  claim 12 , wherein the one or more bioactive agents are selected from the group consisting of macrophage inflammatory protein alpha (MIP-1α), macrophage inflammatory protein beta (MIP-1β), interferon gamma-induced protein 10 (IP-10), interleukin 1 Beta (IL-1β), interleukin 2 (IL-2), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 7 (IL-7), interleukin 8 (IL-8), interleukin 9 (IL-9), interleukin 10 (IL-10), interleukin 12 (IL-12), interleukin 13 (IL-13), interleukin 15 (IL-15), interleukin 17 (IL-17), interleukin 18 (IL-18), interleukin 21 (IL-21), interleukin 22 (IL-22), interleukin 23 (IL-23), interleukin 27 (IL-27), interleukin 31 (IL-31), interferon alpha (INFα), a lymphokine, granulocyte macrophage colony stimulating factor (GM-CSF), stem cell factor (KL), tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), nerve growth factor, platelet-derived growth factor (PDGF), fibroblast growth factor 2 (FGF-2), epidermal growth factor (EGF), keratinocyte growth factor, transforming growth factor (TGF), insulin-like growth factor, des(1-3)-IGF-I (brain IGF-I), neurotrophin-3 (NT-3), brain-derived neurotrophic factor (BDNF), eotaxin, CXC ligand 1 (GROα), interleukin 1 receptor antagonist (IL-1RA), interleukin 1 alpha (IL-1α), leukemia inhibitory factor (LIF), placental growth factor (PLGF), chemokine ligand 5 (RANTES), stem cell factor (SCF), stromal cell derived factor (SDF1α), and tumor necrosis factor (TNF β). 
     
     
         16 . The composition of  claim 1 , wherein the pH of the composition is between 5.0 and 7.4. 
     
     
         17 . The composition of  claim 1 , wherein the pH of the composition is between 4.1 and 7.2. 
     
     
         18 . The composition of  claim 1 , wherein the pH of the composition is between 4.1 and 6.2. 
     
     
         19 . The composition of  claim 1 , further comprising stem cells. 
     
     
         20 . The composition of  claim 1 , wherein the stem cells consist of non-expanded cell populations.