Culturing adult human bone marrow stem cells on gelatin scaffold with pNIPAAm as transplanted grafts for skin regeneration.
Skin tissue engineering is a possible solution for the treatment of extensive skin defect. The ultimate goal of skin tissue engineering is to restore the complete functions of native skin, but until now the structures and functions of skins are only partially restored. By negative immunoselection (CD45 and glycophorin A), we isolated and cultivated adult human bone marrow stem cells (hBMSCs) that are of multilineage differentiation potential. In this study, we first demonstrated that by using gelatin/thermo-sensitive poly N-isopropylacrylamide (pNIPAAm) and the immunocompromised mice model, the hBMSCs possess the differentiation potential of epidermis and the capability of healing skin wounds. The in vitro observations and the results of the scanning electron microscope showed that the hBMSCs can attach and proliferate in the gelatin/thermo-sensitive pNIPAAm. To further monitor the in vivo growth effect of the hBMSCs in the skin-defected nude mice, the green fluorescence protein (GFP) gene was transduced into the hBMSCs by the murine stem cell viral vector. The results showed that the rates of cell growth and wound recovery in the hBMSC-treated group were significantly higher than those in the control group, which was only treated with the gelatin/pNIPAAm (p < 0.01). More importantly, the re-epithelialization markers of human pan-cytokeratin and E-cadherin were significantly increased on day 7, day 14, and day 21 after the hBMSC-scaffold with the pNIPAAM in the mice with skin defects (p < 0.05). Moreover, the stem cell markers of human CD13 and CD105 were gradually decreased during the period of wound healing. In sum, this novel method provides a transferring system for cell therapies and maintains its temperature-sensitive property of easy-peeling by lower-temperature treatment. In addition, the in vitro and in vivo GFP imaging systems provide a new imaging modality for understanding the differentiation process and the effective expression of stem cells in wound healing.
Perng
Cherng-Kang
CK
Department of Surgery, Division of Plastic and Reconstructive Surgery, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan.
Kao
Chung-Lan
CL
Yang
Yi-Ping
YP
Lin
Han-Tso
HT
Lin
Wen-Bin
WB
Chu
Yue-Ru
YR
Wang
Hsiao-Jung
HJ
Ma
Hsu
H
Ku
Hung-Hai
HH
Chiou
Shih-Hwa
SH
D000178:Acrylamides; D000180:Acrylic Resins; D000293:Adolescent; D000328:Adult; D053687:Adult Stem Cells; D000818:Animals; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D018929:Cell Culture Techniques; D005780:Gelatin; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D011108:Polymers; D012038:Regeneration / Q000502:physiology*; D012879:Skin Physiological Phenomena; D033581:Stem Cell Transplantation; D054457:Tissue Scaffolds; D014945:Wound Healing / Q000502:physiology
D000178:Acrylamides; D000180:Acrylic Resins; D011108:Polymers; C052970:poly-N-isopropylacrylamide; D005780:Gelatin