Standardized xeno- and serum-free culture platform enables large-scale expansion of high-quality mesenchymal stem/stromal cells from perinatal and adult tissue sources.

BACKGROUND AIMS: Mesenchymal stem/stromal cells (MSCs) are of interest for the treatment of graft-versus-host disease, autoimmune diseases, osteoarthritis and neurological and cardiovascular diseases. Increasing numbers of clinical trials emphasize the need for standardized manufacturing of these cells. However, many challenges related to diverse isolation and expansion protocols and differences in cell tissue sources exist. As a result, the cell products used in numerous trials vary greatly in characteristics and potency. METHODS: The authors have established a standardized culture platform using xeno- and serum-free commercial media for expansion of MSCs derived from umbilical cord (UC), bone marrow and adipose-derived (AD) and examined their functional characteristics. RESULTS: MSCs from the tested sources stably expanded in vitro and retained their biomarker expression and normal karyotype at early and later passages and after cryopreservation. MSCs were capable of colony formation and successfully differentiated into osteogenic, adipogenic and chondrogenic lineages. Pilot expansion of UC-MSCs and AD-MSCs to clinical scale revealed that the cells met the required quality standard for therapeutic applications. CONCLUSIONS: The authors' data suggest that xeno- and serum-free culture conditions are suitable for large-scale expansion and enable comparative study of MSCs of different origins. This is of importance for therapeutic purposes, especially because of the numerous variations in pre-clinical and clinical protocols for MSC-based products.

A 32-Year-Old Man With Dyspnea and Stridor.

CASE PRESENTATION: A 32-year-old man was admitted to the hospital because of dyspnea on exertion for 2 months. Dyspnea occurred in both inspiration and expiration with an associated wheeze that was more pronounced with exertion. He had no other medical history or allergies. The patient was a current one-pack-a-day smoker for the past 5 years. He denied any close personal contact or recent exposure to any patients with active TB. He denied any current symptoms of chest pain, cough, fever, or changes in weight. On a prior admission for similar symptoms, the patient had been diagnosed with asthma and treated with an inhaled corticosteroid/long-acting beta-agonist with no change or improvement in symptoms.

Human Adipose-Derived Mesenchymal Stromal Cells Exhibit High HLA-DR Levels and Altered Cellular Characteristics under a Xeno-free and Serum-free Condition.

BACKGROUND: We have observed an increased expression of negative markers in some clinical-grade, xeno- and serum-free cultured adipose-derived mesenchymal stem/stromal cell (ADMSC) samples. It gave rise to concern that xeno- and serum-free conditions might have unexpected effects on human ADMSCs. This study aims to test this hypothesis for two xeno- and serum-free media, PowerStem MSC1 media (PS) and StemMACS MSC Expansion Media (SM), that support the in vitro expansion of ADMSCs. METHODS: We investigated the expression of negative markers in 42 clinical-grade ADMSC samples expanded in PS. Next, we cultured ADMSCs from seven donors in PS and SM and examined their growth and colony-forming ability, surface marker expression, differentiation, cell cycle and senescence, as well as genetic stability of two passages representing an early and late passage for therapeutic MSCs. RESULTS: 15 of 42 clinical-grade PS-expanded ADMSC samples showed an increased expression of negative markers ranging from 2.73% to 34.24%, which positively correlated with the age of donors. This rise of negative markers was related to an upregulation of Human Leukocyte Antigen - DR (HLA-DR). In addition, the PS-cultured cells presented decreased growth ability, lower frequencies of cells in S/G2/M phases, and increased ß-galactosidase activity in passage 7 suggesting their senescent feature compared to those grown in SM. Although MSCs of both PS and SM cultures were capable of multilineage differentiation, the PS-cultured cells demonstrated chromosomal abnormalities in passage 7 compared to the normal karyotype of their SM counterparts. CONCLUSIONS: These findings suggest that the SM media is more suitable for the expansion of therapeutic ADMSCs than PS. The study also hints a change of ADMSC features at more advanced passages and with increased donor's age. Thus, it emphasizes the necessity to cover these aspects in the quality control of therapeutic MSC products.

A 69-Year-Old Man With Chronic Cough and Recurrent Pneumonia.

CASE PRESENTATION: A 69-year-old man consulted for a 3-day history of fever, wet cough, and yellow-green phlegm. He denied having any dyspnea, chest pain, hemoptysis, swallowing disorders, choke, chills, asthenia, anorexia, or weight loss. He reported a continuous dry cough and three episodes of pneumonia in the past 4 years. He was a nonsmoker, without any other personal or familial medical history. He had no known professional exposure. He was born and lived in Vietnam but had no known contact with TB in his family or workplace. He was never imprisoned or homeless and did never travel abroad.

A 24-Year-Old Man With Recurrent Hemoptysis.

CASE PRESENTATION: A 24-year-old man was admitted for a new episode of hemoptysis. He reported 3 episodes of hemoptysis in the past 2 years. He had no other medical history and was a nonsmoker. As a Vietnamese person born and living in Vietnam, he was at risk for TB, but had not had contact with those having TB in his family or workplace, was never imprisoned nor homeless, and never traveled abroad. He never experienced pneumonia. He coughed up a small amount of fresh with air red blood (around 5 mL each time) several time for 3 days. He had no dyspnea, no chest pain, no fever, no asthenia, and no anorexia.