The Effect Of HO-1 Gene Modification On The Function Of Placenta-derived Mesenchymal Stem Cells

PART I Effects of heme oxygenase-1 gene on viability of placentalderived mesenchymal stem cellsObjective To study the effects of HO-1 gene on viability of placental-derived mesenchymal stem cells under hypoxia.Method 1.Separate and culture the PMSCs from placenta.2.PMSCs were transfected by lentivirus to express the HO-1 gene stably.3.Exam the proliferation,apoptosis and migration ability of the HO-1 gene engineered PMSCs.Result 1.The PMSCs were Isolated and cultured form placenta successfully.After cultured until the third day,spindle cells can be found and presented clonelike growth.The cells spread to more than 80% of the bottom of the culture bottle after 10 days culture.2.Used the lentivirus-HO-1 to transfer the third-generation cells,the HO-1 gene can be expressed stably.3.HO-1 can decrease the apoptosis of the PMSCs induced by hypoxia,enhance cells' proliferation and migration.Conclusion HO-1 gene has protective effects on the PMSCs under hypoxia.Part II HO-1 participates in regulated PMSCs angiogenesis ability under hypoxia.Objective To investigated the effects of HO-1 gene on the PMSCs hypoxia induced angiogenesis secretion and its effects on the viability of HUVEC.To investigated the expression of angiogenesis-related factors and their effects on HUVECs viability in culture supernatants of HO-1 gene engineered PMSCs,which cultured with hypoxia.Method 1.The m RNA expression of VEGF?PIGF?s Flt-1 and s Eng was measured by real-time PCR.2.The protein expression of VEGF?PIGF?s Flt-1 and s Eng in cultured supernatants was measured by ELISA.3.Using the supernatants from PMSCs cultured under low O2 concentration to treated HUVEC,observed HUVEC proliferation and apoptosis.Using HUVEC-tube formation of PMSCs were tested in vitro HUVEC-tube formation assay.Result 1.The expression of VEGF,PIGF m RNA in HO-1 gene modified PMSCs increased compared with PMSCs.Meanwhile,the expression of s Flt-1,s Eng m RNA was decreased.1.The expression of VEGF in HO-1 gene modified PMSCs cultured supernatants was increased compared with PMSCs.Meanwhile,the expression of s Eng expression decreased.2.Hypoxia treated HO-1 gene modified PMSCs supernatants can enhance the proliferation ability while decreased the number of apoptotic HUVECstube formation were increased by HO-1 over-expression.Conclusion HO-1 gene can increased the PMSCs secretion ability of pro-angiogenic factors under hypoxia conditions.These pro-angiogenic factors can increase the viability of HUVECs.