Joel A. Aronowitz

Adipose stromal vascular fraction isolation: a head-to-head comparison of four commercial cell separation systems.

Background: Supplementation of fat grafts with stromal vascular fraction cells is an emerging technique used to improve graft reliability. A variety of systems for isolating stromal vascular fraction cells are commercially available. The lack of performance data obtained operating the systems in a standardized environment prevents objective assessment of performance. This prospective, blinded study compared performance of four commercially available stromal vascular fraction isolation systems when operated in a clinical outpatient surgery environment. Methods: Four different systems were compared: (1) PNC’s Multi Station, (2) CHA Biotech Cha-Station, (3) Cytori Celution 800/CRS System, and (4) Medi-Khan’s Lipokit with MaxStem. Identical lipoaspirate samples from five separate volunteer donors were used to evaluate system process time, viable cell yield, composition, residual enzyme, and operating costs. Results: The mean processing time ranged from 88 to 115 minutes. The highest mean number of viable nucleated cells was obtained using the Celution System (2.41 × 105 cells/g) followed by the Multi Station (1.07 × 105 cells/g). Lipokit and Cha-Station systems yielded nearly a log fewer nucleated cells (0.35 × 105 cells/g and 0.05 × 105 cells/g, respectively). The Celution System also yielded significantly more endothelial cells, CD34+/CD31− cells, and adipose-derived stem cells (colony-forming unit–fibroblast). Residual enzyme levels observed with the Multi Station, Cha-Station, and Lipokit, respectively, averaged 5.1-, 13.0-, and 57-fold higher than that observed with the Celution System. Conclusions: Although all systems generated measurable amounts of stromal vascular fraction, significant variability exists in the number, identity, and safety profiles of recovered viable cells. Side-by-side clinical trials will be required to establish the relevance of these differences.

Clinical Safety of Stromal Vascular Fraction Separation at the Point of Care.

IntroductionPluripotential cells in adipose tissue may be important in long-term volume retention and regenerative effects of fat grafting. Unfortunately, graft harvest with lipoaspiration significantly depletes the population of stromal vascular cells, which includes adipose stem cells. Stromal vascular fraction (SVF) cells may be isolated from excess lipoaspirate at the point of care and used to replenish fat grafts, a technique termed cell-assisted lipotransfer (CAL). Preclinical and clinical evidence supports the rationale of CAL but clinical adoption of the strategy requires evidence of clinical safety. This prospective, level 1 study reports clinical safety of SVF-enhanced fat grafting using a manual, collagenase-based separation process to isolate autogenous progenitor cells from lipoaspirate at the point of care. MethodsOne hundred sixty-four subjects underwent 174 SVF-enhanced autologous fat grafting procedures at the university stem cell center between August 2009 and November 2014 for a variety of cosmetic and reconstructive indications. ResultsCell-assisted lipotransfer was performed for a variety of cosmetic and reconstructive indications. The mean time of the SVF isolation process was 91 minutes. Because of the frequent concomitant procedures, the average operating room time increased by only 11 minutes. Mean follow-up was 19.9 months. There were no major complications and 6 minor complications. No collagenase or neutral protease related complications were observed. ConclusionsThis series of 174 CAL cases demonstrates that SVF cell isolation using a standardized, manual, collagenase-based process at the POC is equivalent in safety compared to nonenhanced fat grafting. These results support expanded use of CAL in the clinical research setting.

Adipose Stromal Vascular Fraction Isolation: A Head-to-Head Comparison of 4 Cell Separation Systems #2.

IntroductionWith stromal vascular fraction (SVF) cell and adipose-derived stem cell–based technologies translating into the clinical setting, numerous isolation systems have been developed for the point of care isolation of SVF cells from adipose tissue. A relative lack of performance data on these systems can make objective assessment difficult for prospective clinicians. This study compared the performance of 4 SVF cell isolation systems. MethodsFour isolation systems were compared: the MultiStation by PNC International, the LipoKit by MediKhan, the GID SVF-2 platform by GID Europe Ltd, and the StemSource 900/MB system by Cytori Therapeutics, Inc. Identical lipoaspirate samples for 5 separate donors were used. Stromal vascular fraction output was compared in terms of nucleated cell yield, viability, residual collagenase activity, sterility of the output, colony-forming unit–fibroblast frequency, frequency of CD31−/CD34+/CD45− cells, and operating statistics. ResultsMean process time ranged from 65.4 to 120.8 minutes. Mean nucleated cell yield per milliliter of tissue processed ranged from 1.01 × 105 cells/mL to 6.24 × 105 cells/mL. Mean cellular viability ranged from 50.3% to 84.02%. Residual collagenase activity was negligible across all systems. Observed colony-forming unit–fibroblast frequency ranged from 0.495% to 1.704%. No significant difference was observed in frequency of CD31−/CD34+/CD45− cells. Results of the anaerobic/aerobic cultures were mixed. ConclusionsThere was considerable variability between the outputs of each system. The system used by a clinician should be tailored to the individual needs of the practice. There is a range of cost options available. This study may help clinicians make more educated decisions when choosing an isolation system to meet their clinical needs.

Lateral Intercostal Artery Perforator Flap in Breast Reconstruction: A Simplified Pedicle Permits an Expanded Role.

BackgroundThe lateral intercostal artery perforator (LICAP) flap is a versatile second-tier option in breast reconstruction. The flap is rotated from redundant lateral chest fold on an easily dissected skin bridge pedicle without microsurgery in an outpatient setting. This series illustrates safety and effectiveness of the LICAP flap for prosthesis coverage when a muscle flap is not available or desired. In some cases, it even provides adequate soft tissue to reconstruct the breast mound without an implant. MethodsLateral intercostal artery perforator flaps performed for breast reconstruction at an ambulatory surgery center were reviewed. ResultsA total of 39 flaps were performed on an outpatient basis for a variety of breast reconstruction indications. One immediate reconstruction with bilateral LICAP flaps was performed after mastectomy. All remaining flaps were for delayed breast reconstruction. Mean operative time for each flap was 65 minutes, and concomitant procedures were performed in 25 of 27 patients. Follow-up was 5 to 96 months. There was 1 major complication (2.5%) and 5 minor (12.8%) complications. ConclusionsThis series demonstrates unique advantages of the LICAP flap for a variety of breast reconstruction problems, including outpatient setting, no muscle sacrifice, flap reliability, and low donor site morbidity. These results confirm previous reports in post bariatric augmentation that the LICAP flap reliably supplies a large skin/adipose flap from the redundant tissue of the lateral chest fold with minimal morbidity even after radiation. The LICAP flap warrants closer consideration in breast reconstruction.

The effect of fibrin sealant on the healing of laser-resurfaced skin

BACKGROUND Fibrin sealant is an effective hemostatic agent and a useful tissue sealant. Studies have also suggested that fibrin sealant may accelerate the normal wound-healing process. OBJECTIVE This study was designed to ascertain whether fibrin sealant would enhance wound healing after CO(2) laser resurfacing in a guinea pig model. METHODS The CO(2) laser was used to create equal areas of skin resurfacing on both sides of 14 Dunkin Hartley guinea pigs. Fibrin sealant was applied to the treatment side, whereas bacitracin was applied to the control side. Biopsies of these areas were performed on days 1, 3, 7, and 10. A histologic evaluation was performed with the use of a grading scale that compared acute and chronic inflammation, granulation tissue, collagen deposition, and epidermal regeneration. RESULTS The wounds treated with fibrin sealant demonstrated a statistically significant reduction in the degree of acute and chronic inflammation as well as collagen deposition. At day 7, fibrin sealant was noted to enhance neovascularization and result in a slight delay in reepithelialization. All wounds were completely reepithelialized at day 10. No wound infections or other complications were noted as a result of the application of fibrin sealant. CONCLUSIONS Although wound healing was not accelerated, the application of fibrin sealant after CO(2) laser resurfacing diminished the acute and chronic inflammatory response, enhanced neovascularization, and reduced collagen accumulation. Further research is needed to assess whether the effects of fibrin sealant noted in this study result in improved cosmetic healing after CO(2) laser resurfacing. (Aesthetic Surg J 2001;21:509-517.).

A Method for Isolation of Stromal Vascular Fraction Cells in a Clinically Relevant Time Frame

There is increasing interest in the clinical applications of adipose-derived stem cells (ASCs) and the stromal vascular fraction (SVF) based on promising preclinical data. As adipose-derived therapeutics begin to translate into the clinical setting, it is important to maintain patient safety as well as uniformity in technique. Here, we describe a method for isolation of stromal vascular fraction cells in a clinically relevant time frame. Analytical laboratory techniques are mentioned, but respective protocols are not provided here.

Tissue Dissociation Enzymes for Adipose Stromal Vascular Fraction CellIsolation: A Review

With the increasing clinical translation of adipose stromal cell based technologies, it is important to thoroughly report on all aspects of these technologies. In this article, we review the commonly used enzymes for adipose tissue dissociation and stromal vascular fraction isolation. The enzymes reviewed include collagenase, trypsin, clostripain, and the neutral proteases dispase and thermolysin. Enzyme characteristics, the advantages of enzymatic isolation, and other relevant practical and regulatory issues are discussed.

Adipose derived stem cell based therapies or male/female pattern hair loss

In the United States, Alopecia Androgenetica, more commonly known as male or female pattern hair loss, affects approximately 50% of the male population and 45% of the female population [1,2]. Alopecia Androgenetica can be characterized by the gradual thinning of hair emphasized at the crown and frontal scalp, attributed to interactions among various genetic and endocrine factors [3,4]. Individuals impacted by Alopecia Androgenetica are subject to general psychological trauma, as many report a decreased quality of life, lack of self-confidence, and limited social contacts [4-6].