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. A cell suspension for direct application to a patient, said cell suspension produced according to a method comprising the steps of:
(a) subjecting a skin tissue sample comprising dermis and epidermis bonded at a dermal-epidermal junction, to at least a physical and/or chemical dissociating means capable of dissociating cellular stratum in the skin tissue sample for sufficient time to weaken cohesive bonding between tissue stratum;
(b) removing the skin tissue sample from the dissociating means used in step (a) and harvesting, in a reservoir containing a nutrient solution, cells from a dermal surface facing the dermal-epidermal junction and an epidermal surface facing the dermal-epidermal junction, wherein the nutrient solution is (i) free of xenogenic serum, (ii) capable of maintaining viability of the cells until applied to a patient and (iii) suitable for direct application to a recipient region on a patient, thereby to produce a mixed suspension of dermal and epidermal cells; and
(c) filtering the mixed suspension of dermal and epidermal cells to remove large cellular conglomerates greater than 200 μm, to produce a cell suspension that is prepared peri-operatively and is directly applied to the patient;
wherein the method does not comprise the steps of pelleting and resuspending the cells;
wherein the filtered cells are substantially free of cellular stratum and are suspended in the nutrient solution such that the cell suspension is sprayable as airborne droplets.
2. The cell suspension according to claim 1 , wherein the skin tissue sample is autologous to said patient.
3. The cell suspension according to claim 1 , wherein the skin tissue sample provides an expansion ratio of 1:10 to 1:80 in a skin treatment.
4. The cell suspension according to claim 3 , wherein the skin treatment is selected from: epidermal resurfacing; replacement after skin loss; re-pigmentation of an area of skin; treatment of burn wounds, leukoderma, vitiligo, piebaldism, or scars; resurfacing cosmetic dermabrasion; resurfacing after laser treatment; or dermal reconstruction.
5. The cell suspension according to claim 1 , wherein the skin tissue sample is up to 2 cm×2 cm in size.
6. The cell suspension according to claim 1 , wherein the disassociating means comprises an enzyme selected from the group consisting of trypsin, trypsin-EDTA, dispase, collagenase, thermolysin, pronase, hyaluronidase, pancreatin, elastase and papain.
7. The cell suspension according to claim 1 , wherein the nutrient solution is a physiological saline.
8. The cell suspension according to claim 1 , wherein recipient region is receiving tissue grafting.
9. The cell suspension according to claim 1 , wherein recipient region is receiving cell grafting.
10. A cell suspension produced according the method comprising the steps of:
(a) providing a skin tissue sample from a patient, the sample comprising a dermis, an epidermis, and a dermal-epidermal junction therebetween;
(b) harvesting cells from a dermal surface facing the dermal-epidermal junction and an epidermal surface facing the dermal-epidermal junction in a reservoir of nutrient solution thereby to produce a plume of dermal and epidermal cells in the nutrient solution; and
(c) filtering the plume of dermal and epidermal cells to remove cellular conglomerates greater than 200 μm to produce a cell suspension,
wherein the filtered cells are substantially free of cellular stratum and are suspended in the nutrient solution such that the cell suspension is sprayable as airborne droplets, wherein the cell suspension is prepared peri-operatively and directly applied to the patient, and wherein the nutrient solution is free of serum xenogenic to the patient and capable of maintaining viability of the dermal and epidermal cells.
11. The cell suspension according to claim 10 , wherein the skin tissue sample provides an expansion ratio of 1:10 to 1:80 in a skin treatment.
12. The cell suspension according to claim 11 , wherein the skin treatment is selected from: epidermal resurfacing; replacement after skin loss; re-pigmentation of an area of skin; treatment of burn wounds, leukoderma, vitiligo, piebaldism, or scars; resurfacing cosmetic dermabrasion; resurfacing after laser treatment; or dermal reconstruction.
13. The cell suspension according to claim 10 , wherein the skin tissue sample is up to 2 cm×2 cm in size.
14. The cell suspension according to claim 10 , wherein the nutrient solution is a physiological saline.
15. The cell suspension according to claim 10 , wherein recipient region is receiving tissue grafting.
16. The cell suspension according to claim 10 , wherein recipient region is receiving cell grafting.