Improvement in Skin Elasticity Using Red Deer Umbilical Cord Lining Mesenchymal Stem Cell Conditioned Media

Introduction

In a clinical trial, significant improvement in signs of aging was reported after topical application of CALECIM® Professional Multi-Action Cream. CALECIM® is powered by conditioned media derived from Red Deer Umbilical Cord Lining Mesenchymal Stem Cell (RD-CLMSC-CM). RD-CLMSC-CM, also known as PTT-6®, contains a rich mixture of bioactive molecules secreted by stem cells such as peptides, cytokines, growth factors, and other proteins which have been optimized for its skin renewal properties. 

This study aims to understand the mechanism behind these clinical results by investigating how RD-CLMSC-CM influences human dermal fibroblasts (HDFs) – the skin cells responsible for maintaining firmness and elasticity

Method

Elastin and Hyaluronic acid staining

As skin ages, its ability to main hydration and elasticity declines, leading to wrinkles, sagging and loss of firmness. Two key components responsible for keeping the skin resilient are hyaluronic acid (HA) and elastin. 

• HA is a natural substance found in the skin that acts like a sponge, retaining moisture by binding water molecules within the skin
• Elastin is a protein fibre that provides elasticity, allowing the skin to stretch and return to its original shape

Due to their critical roles, our scientists investigated whether treating aged skin cells (HDFs) with RD-CLMSC-CM (PTT-6®) could enhance the expression of HA and elastin. To do so, aged HDFs were cultured under different conditions:

1. Control group: aged HDFs  (skin cells) were cultured in DMEM/10% FCS, a standard cell culture medium
2. Test group: aged HDFs ( skin cells) were cultured in RD-CLMSC-CM (PTT-6®),  which contains bioactive molecules secreted by stem cells

After 48 hours of incubation, the HDFs skin cells) underwent immunocytochemical staining to visualise HA and elastic expression, followed by histochemical staining and photo-colorimetric analysis to quantify the levels of HA and elastin

Cell proliferation study

Another sign of skin aging is slower cell renewal. Scientists tested whether RD-CLMSC-CM (PTT-6®) could boost skin cell proliferation by growing HDFs (aged skin cells) in four different culture media:

• RD-CLMSC-CM (red deer stem cell-derived conditioned media, otherwise known as PTT-6®)
• Human-CLMSC-CM (human-derived stem cell conditioned media)
• Human foreskin fibroblast-CM (known for its high proliferative capacity, making it commonly used as a standard for proliferation studies
• DMEM with 10% FCS (a standard nutrient solution used as control, usually animal derived and NOT umbilical cord)

The cells were allowed to grow for five days, after which they were counted using an automated cell counter to measure how much they have multiplied

Analysis

Skin aging is associated with lower levels of extracellular matrix components such as HA and elastin, resulting in poor skin turgor and elasticity. The function elastin fibres are to provide elasticity, whereas HA retains moisture by binding water molecules within the skin. To see if RD-CLMSC-CM (PTT-6®) could boost the production of these key skin components, we treated skin cells and found that RD-CLMSC-CM (PTT-6®) upregulated HDF (aged skin cells) expression of elastin by 56% and HA by 83% compared with DMEM/10% FCS (control media). Therefore, RD-CLMSC-CM could improve skin elasticity and hydration by stimulating the skin’s own productive of these important molecules

Figure 1. Immunohistochemical staining of elastic and hyaluronic acid in HDF (aged skin cells) in DMEM/10% FCS (control) vs RD-CLMSC-CM (PTT-6®) for 48 hours. The images above show darker staining in the RD-CLMSC-CM-treated cells, indicating a greater production of these components. The bar graph below measures the amount of HA and elastin via optical density 

To further understand the effects of RD-CLMSC-CM (PTT-6®) on the skin, we conducted a cell proliferation study. This study measures how HDFs (aged skin cells) response to different treatments arms. The results showed that RD-CLMSC-CM (PTT-6®) had the strongest effect, increasing HDF proliferation by 113% compared to the control (DMEM/10% FCS). This means that skin cells multiplied more than twice as much with RD-CLMSC-CM. Another similar treatment, H-CLMSC-CM (human stem cell derived conditioned media) had nearly the same effect of 112% increase, while FSF-CM (industry standard animal derived conditioned media, NOT umbilical cord) only led to a 16% increase (Figure 2). This suggests that the proliferative factors in RD-CLMSC-CM (PTT-6®) are as efficacious as those in H-CLMSC-CM (human stem cell-derived conditioned media) and demonstrate cross species efficacy without the toxic effects on HDF (aged skin cells). Therefore, RD-CLMSC-CM is an excellent alternative to H-CLMSC-CM in promoting skin health

Figure 2. Total cell counts were conducted to compare the proliferative effects of DMEM/10% FCS (control), FSF-CM, H-CLMSC-CM and RD-CLMSC-CM

Conclusion

These results demonstrate that RD-CLMSC-CM (PTT-6®) could help the skin stay youthful by (1) stimulating significant production of HA and elastin, (2) being as effective as H-CLMSC-CM (human stem cell derived conditioned media) in exerting proliferative effects on skin cells. Taken together, these findings support the clinical trial results, proving that CALECIM® Professional Multi-Action Cream can visibly improve aging skin.

For the full study, refer to: 
Alhaddad M, Boen M, Wu DC, Goldman MP. Red Deer Umbilical Cord Lining Mesenchymal Stem Cell Extract Cream for Rejuvenation of the Face. J Drugs Dermatol [Internet]. 2019 Apr 1 [cited 2025 Feb 6];18(4):363–6. Available from: https://europepmc.org/article/med/31012565