CELL SUSPENSION PREPARATION TECHNIQUE AND DEVICE
The present invention provides for methods and devices suitable for producing a transplantable cellular suspension of living tissue suitable for grafting to a patient. In applying the method and/or in using the device, donor tissue is harvested, subjected to a cell dissociation treatment, cells suitable for grafting back to a patient are collected and dispersed in a solution that is suitable for immediate dispersion over the recipient graft site.
1 . An apparatus for developing peri-operatively a cell suspension from a skin tissue sample from a patient, and for immediately applying the cell suspension to a graft site of the patient, comprising:
(a) a set of tools for cell harvesting, comprising an enzyme for dissociating cellular stratum in the skin tissue sample and a nutrient solution for cell suspension, wherein the nutrient solution is (i) free of xenogenic serum, (ii) configured to maintain cellular viability and (iii) suitable for direct application to the patient;
(b) a heating means configured to heat an enzyme solution containing the enzyme to a working temperature, wherein upon incubating the skin tissue sample in the heated enzyme solution, the cellular stratum in the skin tissue sample is dissociated;
(c) a reservoir configured to hold the skin tissue sample in fluid containment with the nutrient solution, to permit, following incubation of the skin tissue sample in the heated enzyme solution, manipulation of the skin tissue sample in the reservoir, thereby further separating the cellular stratum and harvesting a plume of cells in the nutrient solution; and
(d) a filter recess comprising a filter means configured to remove large cellular congregates from the plume of cells, thereby forming the cell suspension.
2 . The apparatus of claim 1 , wherein the set of tools further comprises a sterile solution for suspension of the enzyme to provide the enzyme solution.
3 . The apparatus of claim 2 , wherein the apparatus further comprises a first member and a second member, wherein the first member includes (b) the heating means, (d) the filter recess, and (e) a fluid containment well for storage of the nutrient solution; wherein the second member forms (c) the reservoir.
4 . The apparatus of claim 3 , where the heating means is one or more heating pads positioned on top of the first member.
5 . The apparatus of claim 3 , wherein the heating means is housed within a recess in the first member.
6 . The apparatus of claim 5 , wherein the recess further comprises a container located therein into which the skin tissue sample is placed for exposure to the enzyme solution.
7 . The apparatus of claim 1 , wherein the filter recess is configured to receive and hold a tube into which the plume of cells is filtered.
8 . The apparatus of claim 3 , wherein the fluid containment well is a receptacle for a container that holds the nutrient solution.
9 . The apparatus of claim 3 , wherein the reservoir formed by the second member is of sufficient size to permit manipulation of the skin tissue sample.
10 . The apparatus of claim 9 , wherein the first member provides a seat upon which the second member is placed during manipulation of the skin tissue sample.
11 . The apparatus of claim 3 , wherein the first member provides a storage compartment in which the set of tools are stored.
12 . The apparatus of claim 11 , wherein the second member provides a lid to the storage compartment.
13 . The apparatus of claim 12 , wherein an underside of the lid forms the reservoir.
14 . The apparatus of claim 1 , wherein the set of tools include a vessel of the enzyme, a sterile solution for suspension of the enzyme, a sterile nutrient solution, scalpel, forceps, syringe, medicine dropper, cell filter, and/or wound dressings.
15 . The apparatus of claim 1 , further comprising a tool for applying the cell suspension to the graft site.
16 . The apparatus of claim 15 , wherein the tool is a spray nozzle, a pipette, an eye-dropper, or a syringe and needle.
17 . The apparatus of claim 1 , wherein the working temperature is between 30 and 37° C.
18 . The apparatus of claim 17 , wherein the working temperature is between 33 and 37° C.
19 . The apparatus of claim 1 , wherein the heating means is configured to heat the enzyme solution to 37° C. within 2 minutes.
20 . The apparatus of claim 1 , wherein the heating means is configured to maintain the working temperature for at least 45 minutes.