MMT CELL PROCESSING SOLUTIONS

MMT no offers a complete office-based regenerative cell processing solution. The items below are part of the processes meth-odologies we provide as a complete in-office lab solution. Our system gives you the ability to perform on-site processing of adipose tissue (lipoaspirate) and separate regenerative (stromal vascular fraction) cells with proven results. Included in this package are lab supplies, kits, reagents and training that ensures your process and equipment perform at the highest standards possible for maximum viability and concentrations of regenerative cells. We are committed to providing a custom solution that meets the needs of physicians and their patients at an affordable value.

Laminar Flow Clean Bench (left) is specially designed for handling of non-pathogenic biological samples, cells and tissue cultures, controls in microbiology, pharmaceutical products preparation, etc., and are qualified as Class 100 Clean-room Classification.

Incubator (right) is designed for simultaneous heating and mixing of small samples. This unit is comes with interchangeable platforms for stand-ard 15ml and 50ml conical tubes. Maximum capacity is 8 x 50ml or 18 x 15ml. Temperature in the device is adjustable over a broad range, making the unit useful for a variety of applica-tions, including bacterial cultures and digestions. Mechanical convection provides a uniform and stable environment within the chamber.

Centrifuge (right) is the most economical of the universal microprocessor controlled units in the process and accommodates micro-plates and tubes up to 50ml in swing-out and fixed angle rotors. Broad speed range and high g-force make them ideal for applica-tions from clinical to molecular biology. A digital microprocessor controls all operating parameters including speed and time. Over speed conditions are eliminated by the rotor recognition program which automatically identifies each rotor and limits it to the maxi-mum rated speed. Improperly loaded rotors trigger the safety imbalance detection sys-tem which stops operation of the centrifuge. The powerful, induction drive in the Z300 centrifuges quickly accelerates rotors to the preset speed. Two acceleration and deceler-ation rates can be chosen to protect fragile samples. Operation of the centrifuge may be timed, continuous or momentary.

MMT provides optical and imaging solutions (left) on a versatile frame which is ideal for docu-menting cell counts and viability at budget con-scious levels. The long working distance conden-ser (50 mm) and objectives (six supplied) provide brilliant resolved images in both phase contrast and bright field techniques. The condenser swings out to accommodate roller bottles or other large cultivation vessels. A camera or imaging system can be supplied upon request for research and data.

Vortex mixer (right) is a simple device used com-monly in laboratories to mix small vials of liquid. It consists of an electric motor with the drive shaft ori-ented vertically and attached to a cupped rubber piece mounted slightly off-center.

MMT's "Cell Assisted" Processing Service (CAPS)

Adipose-Derived Stromal/Stem Cells (ADSC’s)

Within the stromal vascular fraction there consists a heterogeneous cell population which include circulating blood cells, fibroblasts, pericytes, and endothelial cells as well as adipocyte progenitors; also known as adipose-stromal stem cells (1-3). Direct comparisons between the surface immunophenotype of ADSCs and mesenchymal stem cells are 90% identical. There is a surmountable amount of evidence from both in vitro and in vivo studies that demonstrate the pluripotency of ADSCs from adipose tissue isolated from humans (4). These include adipocytes, chondrocytes, hematopoietic supporting hepatocytes, neuronal-like osteoblasts, and pancreatic and skeletal myocytes.(1-3)

ADSC’s used in Fat Grafting and Sculpting

Some key facts known about ADSCs that can “enhance” cosmetic procedures include:

• They have the potential to secrete angiogenic factors that increase vascular supply and reduce adipocyte apoptosis and fat reabsorption (5)
• ADSC’s react to their micro-environment and thus differentiate along a desired lineage that aids in biomechanical support and function of the re-sculpted area (5)

The Procedure

• Raw lipoaspirate is isolated via liposuction (preferably nutational infrasonic liposculpture)
• Lipoaspirate is washed with phosphate buffered saline (PBS) and centrifuged, creating the first stromal vascular fraction (SVF)
• The layers are separated: The SVF pellet is saved for reinjection, the middle layer is discarded, and the fat is then digested
• Enzyme digestion happens for 15-30 minutes
• Digestion is stopped and then centrifuged to obtain a high-density SVF pellet
• All SVF pellets can be combined to physicians needs for replantation
• On average, our process yields over 10 Billion cells/ml of which over 1% will be pluripotent ADSC’s

Recovery

Based on scientific literature and research on stem cells, including ADSC’s, we can theoretically conclude that (5):

• ADSCs transferred into local tissues secrete cytokines and growth factors that can potentially stimulate recovery
• ADSCs are known to secrete anti-apoptotic factors to maintain tissue health
• ADSCs can help modulate and stimulate the recruitment of endogenous stem cells to the host site
• ADSCs provide antioxidant chemicals, free radical scavengers, and chaperone/heat shock proteins
• ADSCs can possibly deliver new mitochondria to damaged cells to rescue aerobic metabolism

Other Known Benefits

ADSCs that differentiate to produce connective tissue could help to increase production of collagen, elastin, and other extracellular matrix proteins(5) that may improve volume and skin tone.

Stem Cells labeled to detect pluripotency (left) and under a DAPI stain in culture (right)

References:

1. Rodbell M. Metabolism of isolated fat cells. II. The similar effects of phospholipase c (clostridium perfringens alpha toxin) and of insulin on glucose and amino acid metabolism. J Biol Chem. 1966;241:130 –139.

2. Rodbell M. The metabolism of isolated fat cells. IV. Regulation of release of protein by lipolytic hormones and insulin. J Biol Chem. 1966;241:3909 –3917.

3. Rodbell M, Jones AB. Metabolism of isolated fat cells. 3. The similar inhibitory action of phospholipase c (clostridium perfringens alpha toxin) and of insulin on lipolysis stimulated by lipolytic hormones and theophylline. J Biol Chem. 1966;241:140 –142.

4. Annals of Plastic Surgery: April 2010 - Volume 64 – Issue 4 - pp 487-490 PDGF and bFGF Modulate Tube Formation in Adipose Tissue-Derived Stem Cells Keerl, Silke MD; Gehmert, Sebastian MD; Gehmert, Sanga MD; Song, Yao-Hua MD, PhD; Alt, Eckhard MD, PhD

5. Circulation Research: 2007;100;1249-1260 Adipose-Derived Stem Cells for Regenerative Medicine Jeffrey M. Gimble, Adam J. Katz and Bruce A. Bunnell