Background Mesenchymal Stem Cells (MSCs) are multipotent cells with low immuonogenecity, and dynamic tissue repair potential, which explains the overwhelming attention they have attracted in regenerative therapy. One notable challenge in MSCs therapy is the bench to bed timeline of freshly cultured MSCs; it does not exceed 24 h. For use after 24 h, MSC need to be cryopreserved - which can preserve the cells for years - but it is a costly and damaging process. Here we introduce a method to extend the bench to bed lifetime of MSCs up to 4 days without the high cost and cell damaging effects of cryopreservation. Our method is based on preserving the MSCs in human plasma. Methods MSCs of 12 tissue samples - 4 adipose, 4 bone marrow and 4 Wharton’s jelly- were cultured and expanded in standard conditions. Cells harvested from passage 2 or 3 were washed, centrifuged, pelleted, and re-suspended in human plasma. Cell suspensions were refrigerated (5 ± 3 °C) or stored at room temperature (22 ± 3 °C) in a sterile, temperature controlled room. During the next 7 days, two tubes (one from each group) were examined every 24 h to assess MSCs viability and growth potential. On day 3, we assessed MSC cell surface markers and its differentiation potential to adipocyte and osteocyte tissues. Results were analyzed by computing the overall mean and applying the independent-samples t-test to those means. Results The sample means for both cell expansion and cell viability were compared between the two “refrigerator” and “room temperature” groups. Although there was a gradual decrease in cell growth potential between the cells stored for 1 day to those stored for 7 days, we show more than 80% of the cells remain alive for up to 4 days of storage in both groups. The cells reached 80% confluency in under 20 days for all samples stored up to 4 days. No significant differences were observed between the two groups (room temperature and refrigerator stored). The differentiation potential to adipocyte and osteocyte tested on day 3 were positive in all samples. The analysis of cell surface markers tested on day 3 were positive for CD90, CD105, CD73 and negative for CD34, CD45 and HLA-DR. Conclusion We present a method of MSC culture medium using human plasma that can preserve their viability and growth potential for up to 4 days in both room and refrigerator temperatures without losing their stemness characteristics (we recommend use of 5 ± 3 °C). This novel method will allow rapid expansion and therapeutic use of MSCs. Since the cells can be maintained in clinical grade, injection ready state for several days, they can be transported across the globe.

Introduction:One of the most common complications of spinal anesthesia in elective cesarean is a headache, known commonly as post-dural-puncture headache (PDPH). Various methods are mainly recommended such as resting and the use of non-opioid analgesics, caffeine, and codeine, but none of them has been fully effective in its treatment. Hence, this study was conducted to evaluate the effect of magnesium sulfate on the prevention of postdural-puncture headache in the elective cesarean.Method: a total of 68 patients candidate for elective cesarean and admitted to Kamali Hospital were selected using convenient sampling and they were randomly divided into two groups. One group received magnesium and other group received saline. Subjects of case group received magnesium at the dose of 50 mg / kg as bolus and the subjects of control group received normal saline at the same dose as bolus. The incidence of headache and its severity 12, 24, 36, 48 , 60 and 72 hours after surgery were measured in both case and control groups. Results:The mean age of patients in the magnesium sulfate group was 27.94 years with a standard deviation of 5.18 and the mean age of patients in the normal saline group was 29.35 years with a standard deviation of 5.97. The mean body mass index (BMI) in the magnesium sulfate group was 26.34 with a standard deviation of 4.03 and the mean body mass index (BMI) in the normal saline group was 27.15 with a standard deviation of 2.47. Postdural-puncture headache severity was lower in the case group than that in the control group at all times (P <0.05). Conclusion:The results of this study revealed that intravenous administration of magnesium sulfate before elective cesarean in patients undergoing spinal anesthesia significantly decreases the severity of post-duralpuncture headache (PDPH).

Background Mesenchymal Stem Cells (MSCs) are multipotent cells with low immuonogenecity, and dynamic tissue repair potential, which explains the overwhelming attention they have attracted in regenerative therapy. One notable challenge in MSCs therapy is bench to bed timeline of freshly cultured MSCs; it does not exceed 24 hours. For use after 24 hours, MSC need to be cryopreserved, a process that preserves the cells for years, but is costly and damaging. Here we introduce a method to extend the bench to bed lifetime of MSCs up to 4 days without the high cost and cell damaging effects of cryopreservation. Our method uses human plasma as medium. Methods MSCs of 12 tissue samples − 4 adipose, 4 bone marrow and 4 Wharton's jelly- were cultured and expanded under standard conditions. After the cell harvest, each sample was suspended in human plasma. Cell suspensions were refrigerated (5 ± 3° C) or stored at room temperature (22 ± 3°C). During the next 7 days, two tubes (one from each group) were examined every 24 hours to assess MSCs viability and growth potential. On day 3, we assessed MSC 1) differentiation potential to adipocyte and osteocyte tissues 2) surface markers. Results were analyzed by computing the overall mean and applying the independent-samples t-test to those means. Results The sample means for both cell expansion and cell viability were compared between the two “fridge” and “room temperature” groups. Although there was a gradual decrease in cell growth potential between the cells stored for 1 day to those stored for 7 days, we show more than 80% of the cells remain alive for up to 4 days of storage in both groups. The cells reached 80% confluency in under 20 days for all samples stored up to 4 days. No significant differences were observed between the two groups (room temperature and fridge stored). The differentiation potential to adipocyte and osteocyte tested on day 3 were positive in all samples. The analysis of cell surface markers tested on day 3 were positive for CD90, CD105, CD73 and negative for CD34, CD45 and HLA-DR. Conclusion We present a method of MSC culture medium using human plasma that can preserve their viability and growth potential for up to 4 days in both room and refrigerator temperatures without losing their stemness characteristics (we recommend use of 5 ± 3°C). This novel method will allow rapid expansion and therapeutic use of MSCs. Since the cells can be maintained in clinical grade, injection ready state for several days, they can be transported across the globe.