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Study on evaluating Sry as an engrafting track of bone mesenchymal stem cells modified with adenoviral vector expressing human hepatocyte growth factor for healing of burn wounds

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Author:
TA Wei-wei()
HA Xiao-qin()
ZHAO Man(The Second Clinical Medical College, Lanzhou University, Lanzhou 730050, China)
LIU Guo-jun(The Second Clinical Medical College, Lanzhou University, Lanzhou 730050, China)
ZHANG Shang-di()
Journal Title:
China Medicine
Issue:
Volume 06, Issue 12, 2011
DOI:
10.3760/cma.j.issn.1673-4777.2011.12.006
Key Word:
Wound;Bone mesenchymal stem cells;Sty gene;Polymerase chain reaction;DNA probe

Abstract: Objective To prepare digoxin labeled cDNA probe of rat sex determining region of the Y (Sry)and to evaluate the effects of bone mesenchymal stem cells (BMSCs) with transfection of adenoviral vector expressing human hepatocyte growth factor (Ad-HGF) after heterogenous grafting on healing of burn wounds.Methods Sry gene from liver tissue of male rat was cloned by polymerase chain reaction method and constructed to T Easy Vector,prepared in situ hybridization cDNA probe of Sry labeled with Digoxin.BMSCs of male Wistar rat were isolated and cultured through density gradient centrifugation and prepared BMSCs with transfection of Ad-HGF.Sixteen female rats were subjected to boiling water for 12 seconds to produce partial skin bum( pathologic grade Ⅱ ),then injected BMSCs modified with Ad-HGF into underplayed dermis of wound surface.The expressing of Sry gene in histological section through in situ hybridization technique was detected.Results ① Sry gene from 8 male rats was successfully cloned; ②Digoxin labeled cDNA probe of Sty was prepared; ③through in situ hybridization technique,the expression of Sry in cambium tissue of all the 16 female rats' wound surface was positive,while in the normal tissue it was negative.Conclusions Exogenous BMSCs participates repair of wound.Sry can be used as a cell trace maker.This experiment offers reference data for tracing technique of cell therapy and clinical application of BMSCs.

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