Fig 1.
TLRP preparation from turtle blood samples.
Schematic representation (a) and flowchart (b) of the preparation of the Thrombocytes and Leukocytes Rich Plasma (TLRP) from blood sample collected from the caudal or cervical vein of Testudo spp and T. scripta elegans. After a first centrifugation step, blood cells were fractionated on a Histopaque 1.077 gradient and leukocytes/thrombocytes (LL/TT) collected to prepare the TLRP. A step by step summary is available as Supporting information, S5 Fig.
Fig 2.
Blood cells fractionation on Histopaque 1.077 performed applying (a) 480g, 30 min or (b) 150g, 20 min centrifugation step, RT.
Cell counting demonstrated a higher recovery of leukocytes and thrombocytes with method (b).
Table 1.
Clinical cases.
Fig 3.
Upper row: Turtle whole blood cell staining with Diff Quick (a) and PAS (b).
Lower row: TLRP smear stained with Diff Quick (c) and PAS stain (d). Thrombocytes are characterized by a round or oval shape, with a pale cytoplasm positive to PAS stain. (a),(b),(d): 100x magnification. (c): 40x magnification.
Table 2.
Separation of leukocytes and thrombocytes on Hystopaque layer.
Table 3.
Cell-poor plasma and TLRP gelation.
Fig 4.
A severe traumatic injury with bone exposition (radius and ulna, yellow arrows), was referred 60 days after the accident.
TLRP was applied as a gel. After application, the limb was bandaged. Seven days later the lesion was healed, with the production of a protective fibrotic tissue.