Proliferation and differentiation of human adipose-derived mesenchymal stem cells (ASCs) into osteoblastic lineage are passage dependent.

OBJECTIVE: The effect of in vitro expansion of human adipose-derived stem cells (ASCs) on stem cell properties is controversial. We examined serial subcultivation with expansion on the ability of ASCs to grow and differentiate into osteoblastic lineages. DESIGN: Flow cytometric analysis, growth kinetics, cell population doubling time, light microscopy and confocal analysis, and osteogenesis induction were performed to assess growth and osteogenic potential of subcultivated ASCs at passages 2 (P2), P4 and P6. RESULTS: Flow cytometric analysis revealed that ASCs at P2 express classical mesenchymal stem cell markers including CD44, CD73, and CD105, but not CD14, CD19, CD34, CD45, or HLA-DR. Calcium deposition and alkaline phosphatase activity were the highest at P2 but completely abrogated at P4. Increased passage number impaired cell growth; P2 cultures exhibited exponential growth, while cells at P4 and P6 showed near linear growth with cell population doubling times increased from 3.2 at P2 to 4.8 d at P6. Morphologically, cells in various subcultivation stages showed flattened shape at low density but spindle-like structures at confluency as judged by phalloidin staining. CONCLUSIONS: Osteogenic potential of ASCs is impaired by successive passaging and may not serve as a useful clinical source of osteogenic ASCs past P2.

Ultrasound-guided serratus anterior plane block versus thoracic paravertebral block for perioperative analgesia in thoracotomy.

BACKGROUND: Thoracotomy needs adequate powerful postoperative analgesia. This study aims to compare the safety and efficacy of ultrasound (US)-guided serratus anterior plane block (SAPB) and thoracic paravertebral block (TPVB) for perioperative analgesia in cancer patients having lung lobectomy. PATIENTS AND METHODS: This clinical trial involved 90 patients with lung cancer scheduled for lung lobectomy randomly divided into three groups according to the type of preemptive regional block. Group TPVB received US-guided TPVB. In Group SAPB, US-guided SAPB was performed. The patients of the control Group received general anesthesia alone. The outcome measures were postoperative visual analog scale (VAS) score, intraoperative fentanyl consumption, time of first rescue analgesic, total dose postoperative analgesic, and drug-related adverse effects. RESULTS: Analgesia was adequate in TPVB and SAPB groups up to 24 h. VAS score was comparable in TPVB and SAPB groups and significantly lower compared to control group up to 9 h postoperatively. At 12 and 24 h, TPVB group had significantly lower VAS score relative to SAPB and control groups. Total intraoperative fentanyl consumption was significantly lower in TPVB and SAPB Groups compared to control group. The majority of TPVB Group cases did not need rescue morphine, while the majority of control group needed two doses (P < 0.001). The hemodynamic variables were stable in all patients. Few cases reported trivial adverse effects. CONCLUSION: Preemptive TPVB and SAPB provide comparable levels of adequate analgesia for the first 24 h after thoracotomy. TPVB provided better analgesia after 12 h. The two procedures reduce intraoperative fentanyl and postoperative morphine consumption.

Data on isolating mesenchymal stromal cells from human adipose tissue using a collagenase-free method.

The present dataset describes a detailed protocol to isolate mesenchymal cells from human fat without the use of collagenase. Human fat specimen, surgically cleaned from non-fat tissues (e.g., blood vessels) and reduced into smaller fat pieces of around 1-3 mm size, is incubated in complete culture media for five to seven days. Then, cells started to spread out from the fat explants and to grow in cultures according to an exponential pattern. Our data showed that primary mesenchymal cells presenting heterogeneous morphology start to acquire more homogenous fibroblastic-like shape when cultured for longer duration or when subcultured into new flasks. Cell isolation efficiency as well as cell doubling time were also calculated throughout the culturing experimentations and illustrated in a separate figure thereafter. This paper contains data previously considered as an alternative protocol to isolate adipose-derived mesenchymal stem cell published in "Proliferation and differentiation of human adipose-derived mesenchymal stem cells (ASCs) into osteoblastic lineage are passage dependent" [1].